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  • LINC00167 regulates RPE differentiation by targeting the miR-203a-3p/SOCS3 axis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-15
    Xue Chen; Ruxu Sun; Daidi Yang; Chao Jiang; Qinghuai Liu

    Increasing evidences have indicated that long non-coding RNAs (lncRNAs) play significant roles in various diseases; however, their roles in age-related macular degeneration (AMD) remain unclear. Dedifferentiation and dysfunction of retinal pigment epithelium (RPE) cells have been shown to contribute to AMD etiology by several studies. Herein, we found that lncRNA LINC00167 was down-regulated in RPE-choroid samples of AMD patients and dysfunctional RPE cells, and was consistently up-regulated along with RPE differentiation. In vitro study indicated that reduced endogenous LINC00167 expression resulted in RPE dedifferentiation, which was typified by attenuated expression of RPE markers, reduced vascular endothelial growth factor A secretion, accumulation of mitochondrial reactive oxygen species, and interrupted phagocytic ability. Mechanistically, LINC00167 functioned as a sponge for microRNA miR-203a-3p to restore the expression of the suppressor of cytokine signaling (SOCS3), which further inhibited the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. Taken together, our study demonstrated that LINC00167 showed a protective role in AMD by maintaining RPE differentiation through LINC00167/miR-203a-3p/SOCS3 axis and might be a potential therapeutic target for AMD.

    更新日期:2020-01-15
  • 更新日期:2020-01-15
  • Upregulation of long noncoding RNA LINC00460 facilitates gastric cancer progression through epigenetically silencing CCNG2 by EZH2/LSD1 and indicates poor outcomes
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-15
    Jiebin Yang; Yikai Lian; Renzhi Yang; Yifan Lian; Jingtong Wu; Jingjing Liu; Keming Wang; Hongzhi Xu

    Non-protein-coding functional elements in the human genome in postgenomic biology field have been drawing great attention in recent years. Thousands of long noncoding RNAs (lncRNAs) have been found to be expressed in various tumors. Yet only a small proportion of these lncRNAs have been well characterized. We have demonstrated that LINC00460 could affect cell proliferation through epigenetic regulation of KLF2 and CUL4A in human colorectal cancer. However, the clinical significance and biological role of LINC00460 in gastric cancer (GC) remain largely unknown. In this research, we discovered that LINC00460 is remarkably up-regulated in GC tissues compared to the non-tumor tissues. Besides, LINC00460 served as an independent prognostic marker in GC. Functionally, proliferation of GC cells could be regulated by LINC00460 both in vitro and in vivo. RNA-seq analysis for the whole transcriptome indicated that LINC00460 may serve as a key regulatory factor in the tumorigenesis of GC. What’s more, the biological function of LINC00460 was mediated, to certain extent, by the direct interaction with enhancer of zeste homolog 2 (EZH2) and lysine (K)-specific demethylase 1A (LSD1) proteins. Further analyses indicated that LINC00460 promoted GC proliferation at least partly through the down-regulation of tumor suppressor-gene Cyclin G2 (CCNG2) which is mediated by EZH2 and LSD1. In conclusion, our results suggested that LINC00460 acted as an oncogene in GC to inhibit the expression of CCNG2 at least partly by binding with EZH2 and LSD1. Our study could provide additional insights into the development of novel target therapeutic methods for GC.

    更新日期:2020-01-15
  • Novel engineered programmable systems for ADAR-mediated RNA editing
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-15
    Guillermo Aquino-Jarquin

    One of the most prevalent forms of post-transcriptional RNA modification is the conversion of adenosine nucleosides to inosine (A-to-I), mediated by adenosine deaminase acting on RNA (ADAR) enzymes. The advent of the CRISPR/Cas systems inspires researchers to work actively in the engineering of programmable RNA-guided machines for basic research and biomedical applications. In this regard, CIRTS, RESCUE, RESTORE, and LEAPER are innovative RNA base editing platforms that have recently been engineered to perform programmable base conversions on target RNAs mediated by ADAR enzymes, in mammalian cells. Thus, these four currently characterized RNA editing systems constitute novel molecular tools with compelling programmability, specificity, and efficiency that show us some creative ways to take advantage of the engineered deaminases for precise base editing. Moreover, the advanced engineering of these systems permits to edit full-length transcripts containing disease-causing point mutations without the loss of genomic information, providing an attractive alternative for in-vivo research and in the therapeutic setting if the challenges encountered in off-target edits and delivery are appropriately addressed. Here, I present an analytical approach of the current status and rapid progress of the novel ADAR-mediated RNA editing systems when highlighting the qualities of each new RNA editing platform and how these RNA targeting strategies could be used to recruit human ADARs on endogenous transcripts, not only for our understanding of RNA-modification-mediated regulation of gene expression but also for editing clinically relevant mutations in a programmable and straightforward manner.

    更新日期:2020-01-15
  • miR-552 Regulates Liver Tumor-Initiating Cells Expansion and Sorafenib Resistance
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-15
    Tao Han; Yue Zhang; Xiaodan Yang; Lei Han; Hengyu Li; Tingsong Chen; Zhendong Zheng

    MicroRNAs (miRNAs) are involved in tumorigenesis, progression, recurrence and drug resistance of hepatocellular carcinoma (HCC). However, few miRNAs have been identified and entered clinical practice. Herein, we report that miR-552 is upregulated in HCC tissues and has an important function in liver tumor-initiating cells (T-ICs). Functional studies revealed that a forced expression of miR-552 promotes liver T-ICs self-renewal and tumorigenesis. Conversely, miR-552 knockdown inhibits liver T-ICs self-renewal and tumorigenesis. Mechanistically, miR-552 downregulates PTEN via its mRNA 3’UTR and activates AKT phosphorylation. Our clinical investigations elucidated the prognostic value of miR-552 in HCC patients. Furthermore, miR-552 expression determines the responses of hepatoma cells to sorafenib treatment. The analysis of patient cohorts and patient-derived xenografts (PDXs) further demonstrated that miR-552 may predict sorafenib benefits in HCC patients. In conclusion, our findings revealed the crucial role of the miR-552 in liver T-ICs expansion and sorafenib response, rendering miR-552 an optimal target for the prevention and intervention in HCC.

    更新日期:2020-01-15
  • CircRASSF2 acts as competing endogenous RNA and promotes papillary thyroid carcinoma progression through miR-1178/TLR4 signaling pathway
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-15
    Wenhong Zhou; Guojun Wu; Jiyu Li; Xiaona Lin; Yongjie Sun; Hao Xu; Peng Shi; Ling Gao; Xingsong Tian

    Circular RNAs (circRNAs) are a class of non-coding RNAs broadly expressed in cells of various species. However, the molecular mechanisms that link circRNAs with progression of papillary thyroid carcinoma (PTC) are not well understood. In the present study, we attempted to provide novel basis for targeted therapy for PTC from the aspect of circRNA-miRNA-mRNA interaction. We investigated the expression of circRNAs in 5 paired PTC tissues and normal tissues by microarray analysis. The circRNA microarray assay followed by RT-qPCR was used to verify the differential expression of hsa_circ_0059354, which is located on chromosome 20 and derived from RASSF2, and thus we named it circRASSF2. The qRT-PCR analysis was to investigate the expression pattern of circRASSF2 in PTC tissues and cell lines. Then, the effects of circRASSF2 on cell proliferation and apoptosis were assessed in PTC in vitro. Furthermore, bioinformatics online programs predicted and luciferase reporter assay were used to validate the association of circRASSF2 and miR-1178 in PTC cells. In this study, circRASSF2 was observed to be upregulated in PTC tissues and cell lines. Knockdown of circRASSF2 inhibited cell proliferation and promoted cell apoptosis in PTC cells. Bioinformatics analysis predicted that there is a circRASSF2/miR-1178/TLR4 axis in PTC. Dual-luciferase reporter system validated the direct interaction of circRASSF2, miR-1178, and TLR4. Furthermore, circRASSF2 facilitates PTC progression in vivo. Importantly, we demonstrated that circRASSF2 was upregulated in serum exosomes from PTC patients. In summary, our study demonstrated that circRASSF2 modulates PTC progression through miR-1178/TLR4 pathway. Our findings indicated that circRASSF2 may serve as a promising therapeutic target for the treatment of PTC patients.

    更新日期:2020-01-15
  • Nanoparticle delivery of anti-inflammatory locked nucleic acid oligonucleotides prevents airway inflammation in a house dust mite model of asthma
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Sabrina C. Ramelli; Brian S. Comer; Jared M. McLendon; Lydia L. Sandy; Andrew P. Ferretti; Robert Barrington; Jeff Sparks; Majed Matar; Jason Fewell; William T. Gerthoffer
    更新日期:2020-01-15
  • Comparative Analysis of Single-cell Transcriptome Identify Reprogramming Driver Factor for Efficiency Improvement
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Hanshuang Li; Mingmin Song; Wuritu Yang; Pengbo Cao; Lei Zheng; Yongchun Zuo

    Terminally differentiated somatic cells can be reprogrammed into a totipotent state through somatic cell nuclear transfer (SCNT). The incompletely reprogramming is the major reason for developmental arrest of SCNT embryos at early stages. In our studies, we found pathways for autophagy, endocytosis and apoptosis were incompletely activated in NT 2-cell arrest embryos, whereas extensively inhibited pathways for stem cell pluripotency maintenance, DNA repair, cell cycle and autophagy may result in NT 4-cell embryos arrest. As for NT normal embryos, a significant shift in expression of developmental TFs Id1, Pou6f1, Cited1 and Zscan4c were observed. Compared with pluripotent gene Ascl2 only activated in NT 2-cell, Nanog, Dppa2 and Sall4 had major expression waves both in normal development of NT 2-cell and 4-cell embryos. Additionally, Kdm4b/4d and Kdm5b had been confirmed as key markers in NT 2-cell and 4-cell embryos, respectively. Histone acetylase Kat8, Elp6, and Eid1 were co-activated in NT 2 and 4-cell to facilitate embryos normal development. Gadd45a as a key driver functions with Tet1 and Tet2 to improve the efficiency of NT reprogramming. Taken together, our findings provided important theoretical basis for elucidating the potential molecular mechanisms and identified reprogramming driver factor to improve the efficiency of SCNT reprogramming.

    更新日期:2020-01-15
  • The circular RNA circHUWE1 sponges the miR-29b-AKT3 axis to regulate myoblast development
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Binglin Yue; Jian Wang; Wenxiu Ru; Jiyao Wu; Xiukai Cao; Haiyan Yang; Yongzheng Huang; Xianyong Lan; Chuzhao Lei; Bizhi Huang; Hong Chen

    Myogenesis is controlled by a well-established transcriptional hierarchy that coordinates the activities of a set of muscle genes. Rencently, roles in myogenesis have been described for non-coding RNAs, including a role of circular RNA (circRNA) to regulate muscle gene expression. However, the functions of circRNA and the underlying mechanism by which circRNAs affect myogenesis remain poorly understood. In this study, we analyzed circRNA high-throughput sequencing results of bovine skeletal muscle samples, and constructed a circRNA–miRNA–mRNA network according to the competitive endogenous RNA (ceRNA) theory. The putative circHUWE1-miR-29b-AKT3 network was analyzed and its involvement in myogenesis was confirmed through a series of assays. To assess the potential function of this regulation, bovine myoblasts were infected with overexpression plasmids and siRNAs that target circHUWE1. Next, cell proliferation, apoptosis, and differentiation were analyzed using CCK8, EdU, flow cytometry, Western blotting, and RT-qPCR assays. The results suggest that circHUWE1 facilitates bovine myoblast proliferation and inhibits cell apoptosis and differentiation. Next, bioinformatics, dual luciferase reporter assay, and AGO2 RNA immunoprecipitation (RIP) approaches were used to verify the interaction between circHUWE1, miR-29b, and AKT3. Subsequently, we identified that circHUWE1 could directly interfere with the ability of miR-29b to relieve AKT3 suppression, which ultimately activates the AKT signaling pathway. These findings suggested a new regulatory pathway for bovine skeletal muscle development, and also expand our understanding of circRNA functions in mammals.

    更新日期:2020-01-15
  • CRISPR-Cas12a Possess Unconventional DNase Activity that Can Be Inactivated by Synthetic Oligonucleotides
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Bin Li; Jingyue Yan; Youxi Zhang; Wenqing Li; Chunxi Zeng; Weiyu Zhao; Xucheng Hou; Chengxiang Zhang; Yizhou Dong

    CRISPR-Cas12a (CRISPR-Cpf1) was reported to have multiple types of cleavage activities. Without the assistance of CRISPR RNA (crRNA), we investigated DNase activity and substrate specificity of Cas12a orthologs in the presence of diverse divalent metal ions. Cas12a from different species are capable of degrading single-stranded DNA (ssDNA) and (or) double-stranded DNA (dsDNA), depending on the metal ions used. In spite of sharing high sequence similarity and functional domains among diverse Cas12a orthologs, only Acidaminococcus sp. Cas12a (AsCas12a) showed a predominant preference for cleaving ssDNA, whereas no detectable activity toward dsDNA substrate in the presence of magnesium (II) ions. In addition, we found that both AsCas12a and Francisella novicida Cas12a (FnCas12a) caused substantial dsDNA cleavage in the presence of manganese (II) ion. More importantly, the DNase activities can be inhibited by synthetic DNA oligonucleotides with phosphorothioate linkage modifications. Overall, ssDNase activity of the Cas12a orthologs uncovered a distinct approach for DNA cleavage compared to crRNA-guided double stranded DNA breaks, and provided insights into potential biological and therapeutic applications.

    更新日期:2020-01-15
  • The biomarker TCONS_00016233 drives septic AKI by targeting the miR-22-3p/AIFM1 signaling axis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Pan Zhang; Lei Yi; Siyuan Qu; Jinzhong Dai; Xiaozhou Li; Bohao Liu; Huiling Li; Kai Ai; Peilin Zheng; Shuangfa Qiu; Yijian Li; Yinhuai Wang; Xudong Xiang; Xiangping Chai; Zheng Dong; Dongshan Zhang

    The prediction of mortality for septic acute kidney injury (AKI) has been assessed by a number of potential biomarkers, including long non-coding RNAs (lncRNAs). However, the validation of lncRNAs as biomarkers, particularly for the early stages of septic AKI, is still warranted. Our results indicate that the lncRNA TCONS_00016233 is upregulated in plasma of sepsis-associated non-AKI and AKI patients, but a higher cutoff threshold (9.5x105, copy number) provided a sensitivity of 71.9% and specificity of 89.6% for the detection of AKI. The plasma TCONS_00016233 was highly correlated with serum creatinine, TIMP-2, IGFBP7, IL-1β, TNFα, CRP, and urinary TCONS_00016233. LPS induced the expression of lncRNA TCONS_00016233 via TLR4/p38MAPK signal pathway in human renal tubular epithelial (HK-2) cells. Furthermore, TCONS_00016233 mediates the LPS induced HK-2 cells apoptosis and the expression of IL-1β and TNFα. Mechanistically, TCONS_00016233 acts as a ceRNA to prevent miR-22-3p-mediated down-regulation of apoptosis-inducing factor mitochondrion- associated 1(AIFM1). Finally, overexpression of TCONS_00016233 is capable of aggravating the LPS and CLP-induced septic AKI by targeting miR-22-3p/AIFM1 axis. Taken together, our data indicate that TCONS_00016233 may serve as an early diagnosis marker for the septic AKI, possibly acting as a novel therapeutic target for septic AKI.

    更新日期:2020-01-15
  • Binding and structural properties of DNA aptamers with VEGF-A-mimic activity
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Toru Yoshitomi; Misako Hayashi; Takumi Oguro; Keiko Kimura; Fumiya Wayama; Hitoshi Furusho; Keitaro Yoshimoto
    更新日期:2020-01-15
  • CircINSR promotes proliferation and reduces apoptosis of embryonic myoblasts by sponging miR-34a
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-14
    Xuemei Shen; Xiaoyan Zhang; Wenxiu Ru; Yongzhen Huang; Xianyong Lan; Chuzhao Lei; Hong Chen

    As a diverse and abundant class of endogenous RNAs, circular RNAs (circRNAs) participate in processes including cell proliferation and apoptosis. Nevertheless, few researchers have investigated the function of circRNAs in bovine muscle development. Based on existing sequencing data, we identified circINSR. The localization of circINSR in bovine myoblasts was investigated by fluorescence in situ hybridization. Molecular and biochemical assays were used to confirm the role of circINSR in myoblast proliferation and the cell cycle. Mitochondrial membrane potential and annexinⅤ-PE/7-AAD staining assays were performed to assess cell apoptosis. Additionally, interactions between circINSR, miR-34a, and target mRNAs were examined using bioinformatics, luciferase assay, and RNA immunoprecipitation. We found that circINSR was highly expressed in embryonic muscle tissue. Overexpression of circINSR significantly promoted proliferation and reduced apoptosis of embryonic myoblasts. Our data suggested that circINSR may act as a sponge of miR-34a and could function through de-repression of target genes in muscle cells. This study proposes that circINSR may function as a regulator of embryonic muscle development. circINSR regulates cells proliferation and apoptosis through miR-34a-modulated Bcl-2 and CyclinE2 expression.

    更新日期:2020-01-14
  • MicroRNA-122 exerts inhibitory effects on osteoblast proliferation and differentiation in osteoporosis by activating the PCP4-mediated JNK signaling pathway
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-13
    Yi-Chen Meng; Tao Lin; Heng Jiang; Jia Yin; Xiao Ma; Rui Gao; Xu-Hui Zhou

    Osteoporosis is characterized by the reduction of bone mineral density and deterioration of bone quality, which leads to high risk of fractures. Some microRNAs (miRNAs) have been confirmed as potential modulators of osteoblast differentiation to maintain bone mass maintenance. We aimed to clarify whether microRNA-122 (miR-122) could regulate osteoblast differentiation in ovariectomized rats with osteoporosis. miR-122 was upregulated and purkinje cell protein 4 (PCP4) was downregulated in ovariectomized rats. PCP4 was identified as a target of miR-122 by dual-luciferase reporter gene assay. We transfected isolated osteoblasts from ovariectomized rats with miR-122 mimic or inhibitor, or PCP4 overexpression vectors. Proliferation and differentiation of osteoblasts were repressed by the overexpression of miR-122 but enhanced by overexpression of PCP4. miR-122 could induce the activation of the c-Jun NH2-terminal kinase (JNK) signaling pathway, while PCP4 blocked this pathway. Rescue experiments further demonstrated that the inhibiting effects of miR-122 on osteoblast differentiation could be compensated by activation of the PCP4 or inhibition of JNK signaling pathway. Collectively, our data imply that miR-122 inhibits osteoblast proliferation and differentiation in rats with osteoporosis, highlighting a novel therapeutic target for osteoporotic patients.

    更新日期:2020-01-14
  • Upregulation of OIP5-AS1 predicts poor prognosis and contributes to the thyroid cancer cell proliferation and migration through activation of Wnt/β-catenin signaling pathway
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-11
    Qiuli Li; Weichao Chen; Rongzhen Luo; Zhiyi Zhang; Ming Song; Wenkuan Chen; Zhongyuan Yang; Yuanzhong Yang; Zhuming Guo; Ankui Yang

    As a common malignancy, thyroid cancer mainly occurs in endocrine system. There have been accumulating studies on therapeutic methods of thyroid cancer, but its internal molecular mechanism is still not fully understood. LncRNA OIP5-AS1 was confirmed as an oncogene and related to poor prognosis in various cancers. Nevertheless, its role and underlying mechanism remain unclear in thyroid cancer. Here, we observed a significant upregulation of OIP5-AS1 in thyroid cancer tissues and cells and upregulated OIP5-AS1 was correlated with poor prognosis in thyroid cancer. Moreover, OIP5-AS1 knockdown resulted in the inhibited cell proliferation and migration, while overexpressed OIP5-AS1 exhibited the reverse function in thyroid cancer. Besides, OIP5-AS1 was found to positively regulate Wnt/β-catenin signaling pathway. Through mechanism exploration, OIP5-AS1 was discovered to activate Wnt/β-catenin signaling pathway via FXR1/YY1/CTNNB1 axis. Finally, rescue assays indicated that the inhibitive role of silenced OIP5-AS1 in thyroid cancer cell growth and Wnt/β-catenin signaling pathway could be rescued by overexpression of CTNNB1 or addition of LiCl. In conclusion, upregulation of OIP5-AS1 predicted unfavorable prognosis and enhanced thyroid cancer cell growth by activating Wnt/β-catenin signaling pathway.

    更新日期:2020-01-11
  • LncRNA SLC7A11-AS1 promotes chemoresistance by blocking SCFβ-TRCP-mediated degradation of NRF2 in pancreatic cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-11
    Qingzhu Yang; Kai Li; Xuemei Huang; Chen Zhao; Yu Mei; Xinyuan Li; Lin Jiao; Huanjie Yang

    Drug resistance is the major obstacle of gemcitabine-based chemotherapy for the treatment of pancreatic ductal adenocarcinoma (PDAC). Many long non-coding RNAs (lncRNAs) are reported to play vital roles in cancer initiation and progression. Here, we report that lncRNA SLC7A11-AS1 is involved in gemcitabine resistance of PDAC. SLC7A11-AS1 is overexpressed in PDAC tissues and gemcitabine resistant cell lines. Knockdown of SLC7A11-AS1 weakens the PDAC stemness and potentiates the sensitivity of resistant PDAC cells towards gemcitabine in vitro and in vivo. SLC7A11-AS1 promotes chemoresistance through reducing intracellular reactive oxygen species (ROS) by stabilizing NRF2, the key regulator in antioxidant defense. Mechanically, SLC7A11-AS1 is co-localized with β-TRCP1 in the nucleus. The exon 3 of SLC7A11-AS1 interacts with the F-box motif of β-TRCP1, the critical domain that recruits β-TRCP1 to the SCFβ-TRCP E3 complex. This interaction prevents the consequent ubiquitination and proteasomal degradation of NRF2 in the nucleus. Our results demonstrate that the overexpression of SLC7A11-AS1 in gemcitabine resistant PDAC cells can scavenge ROS by blocking SCFβ-TRCP-mediated ubiquitination and degradation of NRF2, leading to low level of intracellular ROS which is required for the maintenance of cancer stemness. These findings suggest SLC7A11-AS1 as a therapeutic target to overcome gemcitabine resistance for PDAC treatment.

    更新日期:2020-01-11
  • MicroRNA134 of ventral hippocampus involves in cocaine extinction-induced anxiety-like and depression-like behaviors in mice
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Yuehan Li; Xue Lu; Jiaxun Nie; Panpan Hu; Feifei Ge; Ti-Fei Yuan; Xiaowei Guan

    We previously found that cocaine abuse could increase microRNA134 (miR134) levels in the hippocampus. Yet, the roles of the miR134 in cocaine-related abnormal psychiatric outcomes remain unknown. In this study, using cocaine-induced conditioned place preference (CPP) mice model, we found that mice exhibit the enhanced anxiety-like and depression-like behaviors during cocaine extinction (CE) period of CPP, accompanied by obviously increased miR134 levels and decreased levels of nineteen genes which are associated with synaptic plasticity, glia activity and neurochemical microenvironments, in the ventral hippocampus (vHP). Knockdown of miR134 in vHP in vivo reversed the changes in fifteen of nineteen potential gene targets of miR134, and rescued the abnormal anxiety-like and depression-like behavioral outcomes in CE mice. In parallel, knockdown of miR134 reversed CE-induced changes in dendritic spines and synaptic proteins, and increased the field EPSP of CA1 pyramidal neurons in the vHP of CE mice. In addition, knockdown of miR134 suppressed the CE-enhanced microglia activity, inflammatory, apoptotic and oxidative stress statuses by in the vHP. Taken together, miR134 may involve in cocaine-associated psychiatric problems, potentially via regulating the expressions of its gene targets that are related to synaptic plasticity and neurochemical microenvironments.

    更新日期:2020-01-11
  • The Potential Therapeutic Role of Exosomal MicroRNA-520b Derived from Normal Fibroblasts in Pancreatic Cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Xiangliang Zhang; Hui Li; Tiantian Zhen; Yu Dong; Xiaojuan Pei; Huijuan Shi

    Pancreatic cancer (PC) remains a major health concern, with conventional cancer treatments exerting little influence on the disease course. MicroRNA-520b (miR-520b) functions as a tumor suppressor in several types of human cancers, while its anti-tumor property in the context of PC is still fundamental. The aim of this study is to identify the potential therapeutic role of miR-520b transferred by exosomes derived from normal fibroblasts (NFs) in PC progression. A gain-of-function study was performed to examine the roles of miR-520b in PC cell line SW1990, which suggested that miR-520b served as a tumor suppressor in PC. In order to confirm the role of exosomal miR-520b, exosomes were isolated from NF culture medium and co-cultured with SW1990 cells. During the co-culture experiments, we disrupted exosome secretion and up-regulated exosomal miR-520b. The in vitro co-culture studies revealed that miR-520b was transferred from NF-derived exosomes to PC cells and thereby suppressed PC cell proliferation, invasion, migration, and stimulated apoptosis. Furthermore, inhibited tumor growth and live metastasis upon elevated miR-520b in exosomes was observed in vivo. Conjointly, our study demonstrates that NF-derived exosomal miR-520b impedes the progression of PC, which contributes to a novel therapeutic role of exosomal miR-520b for treating PC.

    更新日期:2020-01-11
  • A small molecule-controlled Cas9 repressible system
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Youjun Wu; Lu Yang; Tammy Chang; Fouad Kandeel; Jiing-Kuan Yee

    CRISPR-Cas9 has been developed into a powerful molecular tool for genome engineering, and has revolutionized the field of biomedical research. Despite the tremendous potential of CRISPR-Cas9 in biomedical research, precise control of CRISPR-Cas9 over the dose and exposure time is important to expand its applications. In this study, we fused Cas9 with a peptide termed small molecule-assisted shut-off (SMASh) consisting of a protease domain and a degron domain derived from hepatitis C virus (HCV). The presence of SMASh allows tight control of the Cas9 stability via a clinically approved HCV protease inhibitor asunaprevir (ASV). We showed that the engineered Cas9 responded to ASV administration and rapidly degraded in a dose- and time-dependent manner. Cas9 degradation was reversible upon ASV removal that restored the gene editing activity. We also showed that limiting the level of Cas9 in cells increased the specificity of gene editing. The SMASh tag therefore provides an effective tool to control Cas9 stability, allowing an improvement in the accuracy, safety and versatility of the CRISPR-Cas9 system for genome editing and gene regulation studies.

    更新日期:2020-01-11
  • Circular RNA profiling reveals exosomal circ_0006156 as novel biomarker in papillary thyroid cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Guojun Wu; Wenhong Zhou; Xiaohua Pan; Zhigang Sun; Yongjie Sun; Hao Xu; Peng Shi; Jiyu Li; Ling Gao; Xingsong Tian

    Circular RNAs (circRNAs) are a class of non-coding RNAs that are broadly expressed in various biological cells and function in regulating gene expression. However, the molecular mechanisms that link circRNAs with progression of papillary thyroid carcinoma (PTC) are not well understood. In the present study, the function of circ_0006156 (circFNDC3B) was investigated in human PTC cells. First, we detected the expression of circFNDC3B in PTC tissues and PTC cell lines by RT-PCR. A luciferase reporter assay and AGO2-RIP was used to confirm the relationship between circFNDC3B and miR-1178. PTC cells were stably transfected with siRNA against circFNDC3B and cell proliferation, migration and invasion were detected to evaluate the effect of circFNDC3B in PTC, while tumorigenesis was assayed in nude mice. In this study, circFNDC3B was observed to be upregulated in PTC tissues and cell lines. Knockdown of circFNDC3B inhibited cell proliferation and promoted cell apoptosis in PTC cells. Bioinformatics analysis predicted that there is a circFNDC3B/miR-1178/TLR4 axis in PTC. Dual-luciferase reporter system validated the direct interaction of circFNDC3B, miR-1178, and TLR4. Furthermore, circFNDC3B facilitates PTC progression in vivo. Importantly, we demonstrated that circFNDC3B was upregulated in serum exosomes from PTC patients. In summary, our study demonstrated that circFNDC3B modulates PTC progression through miR-1178/TLR4 pathway. Our findings indicated that circFNDC3B may serve as a promising therapeutic target for the treatment of PTC patients.

    更新日期:2020-01-11
  • mmu_circ_0000790 Involves in the Pulmonary Vascular Remodeling in Mice with Hypoxic Pulmonary Hypertension via the microRNA-374c-mediated FOXC1
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Lei Yang; Huan Liang; Xianguo Meng; Li Shen; Zhanjiang Guan; Bingchang Hei; Haitao Yu; Shanshan Qi

    Recently, the identification of several circular RNAs (circRNAs) as vital regulators of microRNAs (miRNAs) underlines the increasing complexity of non-coding RNAs (ncRNAs)-mediated regulatory networks. This study aimed to explore the effects of mmu_circ_0000790 on the biological behaviors of pulmonary artery smooth muscle cells (PASMCs) in hypoxic pulmonary hypertension (HPH). The HPH mouse model and hypoxia-induced PASMC model were initially established, and the expression of mmu_circ_0000790 in the pulmonary vascular tissues and hypoxic PASMCs was determined using RT-qPCR. A series of in vitro experiments such as dual luciferase assay, RNA pull-down and RIP assays were conducted to evaluate the interactions among mmu_circ_0000790, microRNA-374c (miR-374c), and forkhead transcription factor 1 (FOXC1). The potential physiological functions of mmu_circ_0000790, miR-374c and FOXC1 in hypoxic PASMCs were investigated through gain- and loss-of function approaches. Upregulated mmu_circ_0000790 was found in both the HPH-pulmonary vascular tissues and hypoxic PASMCs. Besides, mmu_circ_0000790 could competitively bind to miR-374c and consequently upregulate the target gene of miR-374c, FOXC1. It was also observed that mmu_circ_0000790 induced proliferation and inhibited apoptosis of hypoxic PASMCs, which further promoted the pulmonary vascular remodeling in mice with HPH. Therefore, we speculate that mmu_circ_0000790 may serve as prospective targets for the treatment of patients with HPH.

    更新日期:2020-01-11
  • An Integrated Network-based Discovery of Clinically Approved Formula Fufang-Biejia-Ruangan Pill for Repositioning to Treat Hepatocellular Carcinoma by Inhibiting PI3K/AKT/NF-κB Activation
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Yanqiong ZHANG; Xia MAO; Wenjia CHEN; Xiaodong GUO; Liangxiang YU; Funeng Jiang; Xiaoyue WANG; Weijie LI; Qiuyan GUO; Taixian LI; Na LIN

    Drug repositioning offers new clinical applications for existing drugs with shorter approval processes, less costs and risks than de novo experimental drug development. Fufang-Biejia-Ruangan Pill (FBRP) is the first clinically approved anti-fibrosis herbal formula in China. Whether FBRP could be used to treat hepatocellular carcinoma (HCC) remains unclear. Herein, a total of 161 FBRP candidate targets against HCC were identified according to the topological importance in "hepatic fibrosis-cirrhosis-cancer axis-related gene-FBRP putative target" network, and mostly enriched in PI3K/AKT/NF-κB signaling. Experimentally, FBRP inhibited the liver fibrosis and prevented the development of neoplastic lesions at the early stages of hepatocarcinogenesis in diethylnitrosamine-induced rat HCC model. FBRP inhibited tumor cell proliferation, induced tumor-specific cell death and suppressed tumor progression in HCC rats with preventing the activation of PI3K, AKT and IKΚB proteins, reducing the nuclear accumulation of NFΚB1 protein, and decreasing the downstream proteins' expression. Consistently, FBRP suppressed HCC cell proliferation and induced cell cycle arrest in vitro. Co-treatment of FBRP with PI3K inhibitor exhibited additive inhibitory effect on PI3K/AKT/NF-κB activation. Collectively, our data showed the potentials of FBRP in hepatic fibrosis microenvironment regulation and tumor prevention, suggesting that FBRP may be a promising candidate drug for reduction of fibrogenesis and prevention of HCC.

    更新日期:2020-01-11
  • Hsa_circ_001653 Implicates in the Development of Pancreatic Ductal Adenocarcinoma by Regulating microRNA-377-Mediated HOXC6 Axis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Xiangliang Zhang; Hui Li; Tiantian Zhen; Yu Dong; Xiaojuan Pei; Huijuan Shi

    Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive pancreatic cancer with poor survival rate. Circular RNAs (circRNAs) signatures have been identified in some human cancers, there are little data concerning their presence in PDAC. We investigated the role of hsa_circ_001653, a newly identified circRNA, in the development of PDAC. Hsa-circ-001653 expression was measured in 83 paired normal and tumor tissues surgically resected from PDAC patients. Phenotypic changes of PDAC cells were evaluated by assays for cell viability, cell cycle, invasion, and apoptosis. Tube-like structure formation of human umbilical vein endothelial cells (HUVECs) was examined in the presence of PDAC cells. Cross-talk between hsa_circ_001653 and miR-377/HOXC6 was assessed using dual-luciferase reporter assay, Ago2 immunoprecipitation, and northern blot analysis. Nude mice were inoculated with human PDAC cells for in vivo analysis. Hsa_circ_001653 was an upregulated circRNA in PDAC. Silencing of hsa_circ_001653 in PDAC cells via RNA interference inhibited cell viability, cell-cycle progression, in vitro angiogenesis, and invasive properties, showing pro-apoptotic effect. Hsa_circ_001653 was found to bind to miR-377, which in turn repressed HOXC6 expression. Inhibition of miR-377 by its specific inhibitor restored cell viability, cell-cycle progression, in vitro angiogenesis, and invasive properties in PDAC cells lacking endogenous hsa_circ_001653. When nude mice were inoculated with human PDAC cells, inhibition of hsa_circ_001653 had a therapeutic effect. Collectively, the present study provides an enhanced understanding of hsa_circ_001653 as a therapeutic target for PDAC.

    更新日期:2020-01-11
  • Upregulation of Circular RNA circATRNL1 to Sensitize Oral Squamous Cell Carcinoma to Irradiation
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-10
    Guanhui Chen; Yiming Li; Yi He; Binghui Zeng; Chen Yi; Chao Wang; Xiliu Zhang; Wei Zhao; Dongsheng Yu

    Accumulating evidence has demonstrated that circular RNAs (circRNAs) play important roles in regulating gene expression involved in tumor development. However, the role of circRNAs in modulating the radiosensitivity of oral squamous cell carcinoma (OSCC) and its potential mechanisms have not been documented. We performed high-throughput RNA sequencing (RNA-seq) to investigate the circRNA expression profile in OSCC patients and discovered that the circATRNL1 expression was significantly downregulated and closely related to tumor progression. The circATRNL1 was structurally validated via Sanger sequencing, RNase R treatment, and specific convergent and divergent primer amplification. Importantly, the expression levels of circATRNL1 decreased after irradiation treatment, and upregulation of circATRNL1 enhanced the radiosensitivity of OSCC through suppressing proliferation and the colony survival fraction, inducing apoptosis and cell-cycle arrest. Moreover, we observed that circATRNL1 could directly bind to miR-23a-3p and relieve inhibition for the target gene PTEN. In addition, the tumor radiosensitivity-promoting effect of circATRNL1 overexpression was blocked by miR-23a-3p in OSCC. Further experiments also showed that PTEN can reverse the inhibitory effect of OSCC radiosensitivity triggered by miR-23a-3p. We concluded that circANTRL1 may function as the sponge of miR-23a-3p to promote PTEN expression and eventually contributes to OSCC radiosensitivity enhancement. This study indicates that circANTRL1 may be a novel therapeutic target to improve the efficiency of radiotherapy in OSCC.

    更新日期:2020-01-11
  • Exosomes from mmu_circ_0001359-modified ADSCs attenuate airway remodeling by enhancing FoxO1 signaling-mediated M2-like macrophage activation via sponging miR-183-5p
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-09
    Yan Shang; Yahong Sun; Jing Xu; Xiahui Ge; Zhenli Hu; Jiang Xiao; Yunye Ning; Yuchao Dong; Chong Bai

    Asthma is the most common chronic disease and is characterized by airway remodeling and chronic inflammation. Increasingly, studies have found that the activation and M1 phenotypic transformation of macrophages play important roles in asthma progress, including airway remodeling. However, the reversal of M1 macrophages to the M2 phenotype has been shown to attenuate airway remodeling. Exosomes are nano-sized extracellular vesicles derived from endosomes; they play direct roles in governing physiological and pathological conditions by the intracellular transfer of bioactive cargo such as proteins, enzymes, nucleic acids (miRNA, mRNA, DNA) and metabolites. However, transfer mechanisms are unclear. To uncover potential therapeutic mechanisms, an ovalbumin induced asthma mouse model and lipopolysaccharide induced RAW264.7 macrophages cells were constructed. High-throughput sequencing showed that mmu_circ_0001359 was downregulated in asthmatic mice when compared with normal mice. ADSC-exosome treatment suppressed inflammatory cytokine expression by the conversion of M1 macrophages to the M2 phenotype, under lipopolysaccharide induced conditions. Exosomes from mmu_circ_0001359 overexpression in ADSCs increased therapeutic effects, in terms of cytokine expression, when compared with wild type exosomes. Luciferase reporter assays confirmed that exosomes from mmu_circ_0001359-modified ADSCs attenuated airway remodeling by enhancing FoxO1 signaling mediated M2-like macrophage activation, via sponging miR-183-5p. In conclusion, mmu_circ_0001359-enriched exosomes attenuated airway remodeling by promoting M2-like macrophages.

    更新日期:2020-01-09
  • Effects of miR-34b and miR-892a upregulation and inhibition of ABCB1 and ABCB4 on melatonin-induced apoptosis in vincristine-resistant oral cancer cells
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2020-01-09
    Ming-Ju Hsieh; Chiao-Wen Lin; Shih-Chi Su; Russel J. Reiter; Andy Wei-Ge Chen; Mu-Kuan Chen; Shun-Fa Yang

    Multidrug resistance (MDR) is the resistance of cells toward various drugs commonly used in tumor treatment. The mechanism of drug resistance in oral cancer is not completely understood. Melatonin is an endogenously produced molecule involved in active biological mechanisms including antiproliferation; oncogene expression modulation; antitumor invasion and migration; and anti-inflammatory, antioxidant, and antiangiogenic effects. Despite these functions, the effects of melatonin on vincristine (VCR)-resistant human oral cancer cells remain largely unknown. This study analyzed the role of melatonin in VCR-resistant human oral cancer cells along with the underlying mechanism. We determined that melatonin induced the apoptosis and autophagy of VCR-resistant oral cancer cells; these actions were mediated by AKT, p38, and JNK. Melatonin inhibited ABCB1 and ABCB4 expression in vitro/vivo. Melatonin reduced the drug resistance and promoted the apoptosis of VCR-resistant oral cancer cells through the upregulation of miR-892a and miR-34b-5p expressions. The expression of miR-892a and miR-34b-5p was related to melatonin-induced apoptosis, but not autophagy. Therefore, melatonin is a potential novel chemotherapeutic agent for VCR-resistant human oral cancer cell lines.

    更新日期:2020-01-09
  • RNA secondary structure motifs of the influenza A virus as targets for siRNA-mediated RNA interference
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Julita Piasecka; Elzbieta Lenartowicz; Marta Soszynska-Jozwiak; Barbara Szutkowska; Ryszard Kierzek; Elzbieta Kierzek

    The influenza A virus is a human pathogen that poses a serious public health threat due to rapid antigen changes and emergence of new, highly pathogenic strains with potential to become easily transmitted in the human population. The viral genome is encoded by eight RNA segments, and all stages of the replication cycle are dependent on RNA. In this study, we designed siRNA targeting influenza segment 5 NP mRNA structural motifs that encode important functions. The new criteria for choosing the siRNA target was the prediction of accessible regions based on the secondary structure of segment 5 (+)RNA. This design led to siRNAs that significantly inhibit influenza virus type A replication in MDCK cells. Additionally, chemical modifications with the potential to improve siRNA properties were introduced and systematically validated in MDCK cells against the virus. A substantial and maximum inhibitory effect was achieved at concentrations as low as 8 nM. The inhibition of viral replication reached approximately 90% for the best siRNA variants. Additionally, selected siRNAs were compared with antisense oligonucleotides targeting the same regions; this revealed that effectiveness depends on both the target accessibility and oligonucleotide antiviral strategy. Our new approach of target-site preselection based on segment 5 (+)RNA secondary structure led to effective viral inhibition and a better understanding of the impact of RNA structural motifs on the influenza replication cycle.

    更新日期:2019-12-25
  • Long non-coding RNA LOXL1-AS1 drives breast cancer invasion and metastasis by antagonizing miR-708-5p expression and activity
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Huiting Dong; Qun Liu; Tingting Zhao; Fan Yao; Yingying Xu; Bo Chen; Yunfei Wu; Xinyu Zheng; Feng Jin; Jiguang Li; Peng Xing

    LOXL1-AS1, a recently characterized long non-coding RNA (lncRNA), has been reported to modulate tumor progression in several types of cancer. However, the expression and role of LOXL1-AS1 in breast cancer remains unclear. In this study, we sought to identify novel lncRNA regulators engaged in breast cancer metastasis. To this end, we examined 42 cancer-related lncRNAs between MCF7 (with low metastatic potential) and MDA-MB-231 (with high metastatic potential) cells. These lncRNAs have been found to affect the invasiveness of several cancer types, but are still undefined in breast cancer. Among the 42 candidates, LOXL1-AS1 is significantly increased in MDA-MB-231 cells relative to MCF7 cells. We also show that LOXL1-AS1 is upregulated in breast cancer tissues and cells compared to noncancerous counterparts. Increased LOXL1-AS1 expression is correlated with tumor stage and lymph node metastasis in breast cancer patients. Biologically, overexpression of LOXL1-AS1 enhances and knockdown of LOXL1-AS1 suppresses breast cancer cell migration and invasion. In vivo studies demonstrate that depletion of LOXL1-AS1 inhibits breast cancer metastasis. Mechanistically, LOXL1-AS1 sponges miR-708-5p to increase NF-κB activity. LOXL1-AS1 can also interact with EZH2 protein to enhance EZH2-mediated transcriptional repression of miR-708-5p. Rescue experiments indicate that co-expression of miR-708-5p attenuates LOXL1-AS1-induced invasiveness in breast cancer. In addition, there is a negative correlation between LOXL1-AS1 and miR-708-5p expression in breast cancer specimens. Overall, LOXL1-AS1 upregulation facilitates breast cancer invasion and metastasis by blocking miR-708-5p expression and activity. LOXL1-AS1 serves as a potential therapeutic target for breast cancer treatment.

    更新日期:2019-12-25
  • 更新日期:2019-12-25
  • Correction of the aprt gene using repair-PolyPurine Reverse Hoogsteen hairpins in mammalian cells
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Alex J. Félix; Carlos J. Ciudad; Véronique Noé
    更新日期:2019-12-25
  • Melatonin regulates breast cancer progression by the lnc010561/miR-30/FKBP3 axis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Peng Liu; Xinhua Xie; Anli Yang; Yanan Kong; Diane Allen-Gipson; Zhi Tian; Liye Zhou; Hailin Tang; Xiaoming Xie

    Melatonin have been recognized to slow breast cancer growth. The molecular mechanisms may involve long non-coding RNAs (lncRNAs). However, little is known on how melatonin affects lncRNAs expression and function in breast cancer. We used microarrays to explore the expression profile of mRNAs and lncRNAs in melatonin-treated breast cancer cells. KEGG and Reactome pathways analysis were performed to identify the signaling pathways affected by altered expressed mRNAs after melatonin treatment. To explore the functions and mechanisms of the selected differentially expressed mRNA and lncRNA in breast cancer, we performed a series of experiments. We found that FKBP3 and lnc010561 were down-regulated in melatonin-treated breast cancer cells. Knockdown of FKBP3 and lnc010561 inhibited breast cancer proliferation and invasion, and induced apoptosis. And lnc010561 and FKBP3 functioned as competing endogenous RNAs (ceRNAs) for miR-30. Our findings suggested that melatonin regulated breast cancer progression by the lnc010561/miR-30/FKBP3 axis. Melatonin may therefore function as an anticancer strategy for breast cancer.

    更新日期:2019-12-25
  • A Universal Surrogate Reporter for Efficient Enrichment of CRISPR/Cas9-Mediated Homology-Directed Repair in Mammalian Cells
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Nana Yan; Yongsen Sun; Yuanyuan Fang; Jingrong Deng; Lu Mu; Kun Xu; Joe S. Mymryk; Zhiying Zhang

    CRISPR/Cas9-mediated homology-directed repair (HDR) can be leveraged to precisely engineer mammalian genomes. However, the inherently low efficiency of HDR often hampers to identify the desired modified cells. Here, we developed a novel Universal Surrogate Reporter system that efficiently enriches for genetically modified cells arising from CRISPR/Cas9-induced HDR events (namely, HDR-USR). This episomally based reporter can be self-cleaved and self-repaired via HDR to create a functional puromycin selection cassette without compromising genome integrity. Co-transfection of the HDR-USR into host cells and transient puromycin selection achieves efficiently enrichment of HDR-modified cells. We tested the system for precision point mutation at 16 loci in different human cell lines and one locus in two rodent cell lines. This system exhibited dramatic improvements in HDR efficiency at single locus (up to 20.7-fold), two loci at once (42% editing efficiency compared to zero in the control), as well as greatly improved knock-in efficiency (8.9-fold) and biallelic deletion (35.9-fold) at test loci. Further increases were achieved by co-expression of yeast Rad52 and linear single-/double-stranded DNA donor. Taken together, our HDR-USR system provides a simple, robust and efficient surrogate reporter for enrichment of CRISPR/Cas9 induced HDR-based precision genome editing across various targeting loci in different cell lines.

    更新日期:2019-12-25
  • miR-144-3p suppresses osteogenic differentiation of bone marrow mesenchymal stem cells from patients with aplastic anemia through repression of TET2
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Ning Li; Lina Liu; Yuzhang Liu; Suxia Luo; Yongping Song; Baijun Fang

    Reduced osteogenic capacity of bone marrow mesenchymal stem cells (BMSCs) has been causally linked to the development of aplastic anemia. In this work, we aimed to identify novel microRNAs (miRs) that participate in the regulation of differentiation of BMSCs from patients with aplastic anemia. We show that miR-144-3p is significantly upregulated in BMSCs from patients with aplastic anemia relative to control equivalents. Depletion of miR-144-3p significantly enhances osteogenic differentiation of BMSCs from patients with aplastic anemia after culturing in osteogenesis-inducing medium. Conversely, overexpression of miR-144-3p blocks osteogenic differentiation of BMSCs. Mechanistically, miR-144-3p negatively regulates the expression of TET2 in BMSCs. Reduced TET2 expression is associated with a significant decrease in global 5-hydroxymethyl-cytosine levels and osteogenic gene expression. Knockdown of miR-144-3p elevates the expression of TET2 and total 5hmC levels in BMSCs. Silencing of TET2 inhibits the osteogenic differentiation of BMSCs. Overexpression of TET2 reverses miR-144-3p-mediated inhibition of osteogenesis. In addition, there is a significant negative correlation between the expression of miR-144-3p and TET2 in BMSCs from patients with aplastic anemia. Overall, miR-144-3p impairs osteogenic capacity of BMSCs from patients with aplastic anemia through repression of TET2. Therefore, targeting miR-144-3p may be a therapeutic strategy against aplastic anemia.

    更新日期:2019-12-25
  • Circulating lncRNA UCA1 promotes malignancy of colorectal cancer via the miR-143/MYO6 axis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-24
    Yunpeng Luan; Xiang Li; Yunqi Luan; Rong Zhao; Yanmei Li; Lili Liu; Yizhuo Hao; Burakovaov Oleg.Vladimir; Lu Jia

    Exosomes mediate cell-cell crosstalk in cancer progression by transferring a variety of biomolecules, including long noncoding RNAs (lncRNAs). Long non-coding RNA urothelial carcinoma-associated (UCA1) is a well-known lncRNA associated with the development and progression of various cancers, including colorectal cancer (CRC). However, the presence of UCA1 in exosomes and the roles and clinical values of exosomal UCA1 in CRC remain unknown. In this study, we systematically analyzed the expression profiles of exosomal lncRNAs in CRC patients using a high-throughput microarray assay. Then, we evaluated the UCA1 expression levels in a series of CRC tissues and serum exosome of CRC patients using qRT-PCR. The roles of UCA1 on CRC in vitro and in vivo were investigated by MTT, colony formation, transwell, RT-qPCR, flow cytometry, and Western blotting. The miRNA binding sites of UCA1 was predicted using miRcode online website and miR-143 was validated to target UCA1 by dual-luciferase activity assay and AGO2 RNA immunoprecipitation. Finally, the role of exosome-mediated UCA1 was further investigated by co-culturing with CRC cells. This study showed that UCA1 was upregulated in CRC tissues and functioned as an oncogene in CRC. Loss-of-function investigations showed that inhibition of UCA1 suppressed CRC cell proliferation and metastasis in vivo and in vitro. Mechanistically, UCA1 was identified as a miR-143 sponge. We also found that MYO6 was a direct target of miR-1205, which functioned as an oncogene in CRC. Moreover, UCA was also up-regulated in serum exosome of CRC patients and could transfer UCA1 to CRC cells to increase their abilities of cell proliferation and migration. In conclusion, these data suggest that UCA1 could be an oncogene for CRC, and may be served as a candidate target for new therapies in human CRC.

    更新日期:2019-12-25
  • N6-Methyladenosine (m6A): a potential breakthrough for human cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-20
    Lina Liu; Yuwei Wang; Jie Wu; Jingwen Liu; Zongchang Qin; Hong Fan

    Among more than 100 types of identified RNA modification, N6-methyladenosine (m6A) modification is the predominant mRNA modification, which regulates RNA splicing, translocation, stability and translation. m6A modification plays critical roles in the growth, differentiation and metabolism of cells. As a dynamic and reversible modification, m6A is catalyzed by ‘writers’ (RNA methyltransferases), removed by ‘erasers’ (demethylases), and interacts with ‘readers’ (m6A-binding proteins). With more advanced technology applied to researches, molecular mechanisms of RNA methyltransferase, demethylase and m6A-binding protein have been revealed. An increasing number of researches have implicated the correlation between m6A modification and human cancers. Here, we summarize that the occurrence and development of various human cancers are associated with aberrant m6A modification. We also discuss the progress in researches related to m6A modification, providing novel therapeutic insight and potential breakthrough in anticancer therapy.

    更新日期:2019-12-20
  • Circular RNA circFOXM1 plays a role in papillary thyroid carcinoma by sponging miR-1179 and regulating HMGB1 expression
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-20
    Mao Ye; Haitao Hou; Minghai Shen; Shu Dong; Tao Zhang

    Circular RNAs (circRNAs) are a class of non-coding RNAs broadly expressed in cells of various species. However, the molecular mechanisms that link circRNAs with progression of papillary thyroid carcinoma (PTC) are not well understood. In the present study, we attempted to provide novel basis for targeted therapy for PTC from the aspect of circRNA-miRNA-mRNA interaction. We investigated the expression of circRNAs in 5 paired PTC tissues and normal tissues by microarray analysis. The circRNA microarray assay followed by RT-qPCR were used to verify the differential expression of circFOXM1 (hsa_circ_0025033), which located at chr12: 2966846-2983691 and derived from FOXM1. The spliced length of circFOXM1 is 3410 nt. The qRT-PCR analysis was to investigate the expression pattern of circ-FOXM1 in PTC tissues and cell lines. Then, the effects of circ-FOXM1 on tumor growth were assessed in PTC in vitro and in vivo. Furthermore, bioinformatics online programs predicted and luciferase reporter assay were used to validate the association of circFOXM1 and miR-1179 in PTC cells. In this study, circ-FOXM1 was observed to be upregulated in PTC tissues tissues and cell lines. CircFOXM1 down-regulation inhibited tumor growth of PTC in vitro and in vivo. Bioinformatics analysis predicted that there is a circ-FOXM1/miR-1179/HMGB1 axis in PTC. Dual-luciferase reporter system validated the direct interaction of circFOXM1, miR-1179, and HMGB1. In summary, our study demonstrated that circFOXM1 modulates cancer progression through miR-1179/HMGB1 pathway in PTC. Our findings indicate that circFOXM1 may serve as a promising therapeutic target for the treatment of PTC patients.

    更新日期:2019-12-20
  • Chemical diversity of locked nucleic acid-modified antisense oligonucleotides allows optimization of pharmaceutical properties
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-18
    Natalia Papargyri; Malene Pontoppidan; Mikael R. Andersen; Troels Koch; Peter H. Hagedorn

    The identification of molecules that can modulate RNA or protein function, and the subsequent chemical and structural optimization to refine such molecules into drugs, is a key activity in drug discovery. Here we explored the extent to which chemical and structural differences in antisense oligonucleotides, designed as gapmers and capable of recruiting RNase H for target RNA cleavage, can affect their functional properties. To facilitate structure-activity learning, we analyzed two sets of iso-sequential locked nucleic acid (LNA)-modified gapmers, where we systematically varied the number and positions of LNA modifications in the flanks. In total, we evaluated 768 different and architecturally diverse gapmers in HeLa cells for target knockdown activity and cytotoxic potential and found widespread differences in both of these properties. Binding affinity between gapmer and RNA target, as well as the presence of certain short sequence motifs in the gap region, can explain these differences, and we propose statistical and machine learning models that can be used to predict region-specific, optimal LNA-modification architectures. Once accessible regions in the target of interest has been identified, our results show how to refine and optimize LNA-gapmers with improved pharmacological profiles targeting such regions.

    更新日期:2019-12-19
  • Endogenous microRNA competition as a mechanism of shRNA-induced cardiotoxicity
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-18
    Meredith M. Course; Kathryn Gudsnuk; Nitin Desai; Joel R. Chamberlain; Paul N. Valdmanis
    更新日期:2019-12-19
  • 更新日期:2019-12-19
  • DBMDA: A unified embedding for sequence-based miRNA similarity measure with applications to predict and validate MicroRNA-disease associations
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-18
    Kai Zheng; Zhu-Hong You; Lei Wang; Yong Zhou; Li-Ping Li; Zheng-Wei Li

    MicroRNAs (miRNAs) play a critical role in human diseases. Determining the association between miRNAs and disease contributes to elucidate the pathogenesis of liver diseases and seek the effective the treatment method. Despite great recent advances in the field of the associations between miRNAs and diseases, implementing association verification and recognition efficiently at scale presents serious challenges to biological experimental approaches. Thus, computational methods for predicting miRNA–disease association become a research hot spot. In this paper we present a new computational method, named Distance-Based sequence similarity for MiRNA–Disease Association prediction (DBMDA), that directly learns a mapping from miRNA sequence to a Euclidean space. The notable feature of our approach consists in inferring global similarity from region distances which can be figured by chaos game representation algorithm based on the miRNA sequences. In the 5-fold cross-validation experiment, the AUC obtained by DBMDA in predicting potential miRNA-disease associations reached 0.9129. To assess the effectiveness of DBMDA more effectively, we compared it with different classifiers and former prediction models. Besides, we constructed two case studies for Prostate Neoplasms and Colon Neoplasms. Results show that 39 and 39 out of the top 40 predicted miRNAs were confirmed by other databases, respectively. BDMDA has made new attempts in sequence similarity and achieved excellent results, while at the same time providing a new perspective for predicting the relationship between diseases and miRNAs. The source code and datasets explored in this work are available at http://220.171.34.3:81/.

    更新日期:2019-12-19
  • The Long Noncoding RNA HOTTIP Is Highly Expressed in Colorectal Cancer and enhances cell proliferation and invasion
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-18
    Tao Liu; Helei Wang; Haiyao Yu; Miaomiao Bi; Zhenkun Yan; Sen Hong; Shiquan Li

    Long non-coding RNAs (lncRNAs) are associated with a spectrum of biological processes such as gene regulation on transcriptional and post-transcriptional levels. The HOXA transcript at the distal tip (HOTTIP) lncRNA plays an important role in carcinogenesis, however, the underlying role of HOTTIP in colorectal carcinoma (CRC) remain unknown. The aim of the present study was to evaluate the expression and function of HOTTIP in CRC. In the present study, we analyzed HOTTIP expression levels of CRC patients in tumor and adjacent normal tissue by real-time quantitative PCR. Knockdown of HOTTIP by RNA interference was performed to explore its roles in cell proliferation, migration and invasion. Our results found that HOTTIP was up-regulated in human primary CRC tissues. Knockdown of HOTTIP inhibited CRC cell proliferation, migration and invasion. Above all, knockdown of HOTTIP could represent a rational therapeutic strategy for CRC.

    更新日期:2019-12-19
  • circCELSR1 (hsa_circ_0063809) contributes to paclitaxel resistance of ovarian cancer cells by regulating FOXR2 expression via miR-1252
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-18
    Shu Zhang; Jie Cheng; Chenlian Quan; Hao Wen; Zheng Feng; Qin Hu; Jun Zhu; Yan Huang; Xiaohua Wu

    Ovarian cancer is the malignant tumor of the female reproductive system with the highest fatality rate. Tolerance of chemotherapeutic drugs like paclitaxel (PTX) occurring in very early stage is one of the important factors of the poor prognosis of ovarian cancer. Here we aim to study the dysregulation of a particular circular RNA (circRNA), circCELSR1 (hsa_circ_0063809), and its role in in progression and PTX resistance of ovarian cancer. The high expression of circCELSR1 in PTX resistant tissues of ovarian cancer and PTX-resistant ovarian cancer cells (SKOV3/PTX and HeyA-8/PTX) were determined by microarray analyses and Real-Time qPCR. CCK8 assay were performed to investigate the effect of circCELSR1 on PTX sensitivity of ovarian cancer cells. Flow cytometer assays were used to detect cell cycle and apoptosis of ovarian cancer cells. The effect of circCELSR1 on ovarian cancer cells was assessed in vitro and in vivo. The microRNA (miRNA) sponge mechanism of circRNAs was demonstrated using dual-luciferase reporter and RNA immunoprecipitation assays. By microarray (5 PTX-resistant ovarian cancer tissues νs. 5 PTX-sensitive ovarian cancer tissues) and RT- qPCR (36 normal ovarian tissues and ovarian cancer tissues) we identified circCELSR1 to be dramatically high expressed in ovarian cancer samples and correlated with PTX resistance. Compared with sensitive cell lines, circCELSR1 was also high expressed in PTX resistant ovarian cancer cell lines, and circCELSR1 silencing enhanced PTX-induced cytotoxicity in ovarian cancer cells. Meanwhile the inhibition of circCELSR1 also caused ovarian cancer cells G0/G1 arrest and apoptosis increase. In vivo studies revealed that circCELSR1 was stably inhibited in a xenograft mouse model and inhibited the growth of ovarian cancer. Furthermore, we demonstrated that circCELSR1 acts as a sponge for miR-1252 and verified that Forkhead box 2 (FOXR2) is a novel target of miR-1252. Here we explored the specific mechanisms of PTX resistance and tumor progress of ovarian cancer due to circCELSR1, presented the circCELSR1-miR-1252-FOXR2 axis and its role in ovarian cancer drug-sensitivity and progression, and the results may provide experimental basis for clinical application.

    更新日期:2019-12-18
  • Long Non-coding RNA LINC00460 Silencing Represses Epithelial-Mesenchymal Transition in Colon Cancer through Downregulation of ANXA2 via Upregulating miR-433-3p
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-18
    Weiwen Hong; Hongan Ying; Feng Lin; Ruliang Ding; Weiya Wang; Meng Zhang

    Colon cancer (CC), one of the major causes of tumor-associated death, is often presented with a heterogenic pool of cells with unique differentiation patterns. This study explored the functions that LINC00460 displayed in CC by regulating microRNA-433-3p (miR-433-3p) and ANXA2. LINC00460 expression was either silenced or overexpressed in HCT-116 and LOVO cells to explore the functional roles of LINC00460 in CC. The relationship between miR-433-3p and LINC00460/ANXA2 was analyzed using dual-luciferase reporter assay, RNA-pull down, and RIP assays. Cell proliferation, metastasis, invasion, and apoptosis were examined in vitro, and tumorigenicity was evaluated in vivo following LINC00460 silencing. Additionally, the regulatory mechanisms were investigated using LINC00460 and ANXA2 gain- or loss-of-function experiments. We found that LINC00460 was expressed highly in CC. Down-regulation of LINC00460 inhibited cell invasion and proliferation in vitro and restrained tumor growth in vivo. Moreover, LINC00460 was able to specifically bind to miR-433-3p to increase the expression of ANXA2. Furthermore, LINC00460 upregulated the E-cadherin expression and downregulated the vimentin and N-cadherin expression by upregulating ANXA2, therefore inducing epithelial-mesenchymal transition. These findings suggested that LINC00460 might function as an oncogenic lncRNA in CC development and could be explored as a potential biomarker and therapeutic target for CC.

    更新日期:2019-12-18
  • Improved Nucleic Acid Therapy with Advanced Nanoscale Biotechnology
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-17
    Yuhua Weng; Qianqian Huang; Chunhui Li; Yongfeng Yang; Xiaoxia Wang; Jie Yu; Yuanyu Huang; Xing-Jie Liang

    Due to a series of systemic and intracellular obstacles in nucleic acid (NA) therapy, including fast degradation in blood, renal clearance, poor cellular uptake, and inefficient endosomal escape, NAs may need delivery methods to transport to the cell nucleus or cytosol to be effective. Advanced nanoscale biotechnology-associated strategies, such as controlling the particle size, charge, drug loading, response to environmental signals or other physical/chemical properties of delivery carriers, have provided great help for the in vivo and in vitro delivery of NA therapeutics. In this review, we introduce the characteristics of different NA modalities and illustrate how advanced nanoscale biotechnology assist NA therapy. The specific features and challenges of various nanocarriers in clinical and preclinical studies are summarized and discussed. With the help of advanced nanoscale biotechnology, some of the major barriers to the development of NA therapy will eventually be overcome in the near future.

    更新日期:2019-12-18
  • The mesenchymal stem cell-derived exosomal long noncoding RNA H19 promotes wound healing in diabetic foot ulcers by up-regulating PTEN via microRNA-152-3p
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-14
    Bo Li; Song Luan; Jing Chen; Yue Zhou; Tingting Wang; Zhijuan Li; Yili Fu; Aixia Zhai; Changlong Bi

    Mesenchymal stem cells (MSCs) have been reported to hold promise to accelerate wound-healing process in diabetic foot ulcer (DFU) due to the multi-lineage differentiation potential. Hence, this study intended to explore the wound healing role of MSCs-derived exosomes containing long noncoding RNA (lncRNA) H19 in DFU. LncRNA H19 was predicated to bind to microRNA-152-3p (miR-152-3p), which targeted PTEN. Fibroblasts in DFU samples exhibited highly expressed miR-152-3p and poorly expressed lncRNA H19 and PTEN, along with activated PI3K/AKT signaling pathway. The fibroblasts were co-cultured with lncRNA H19-transfected MSCs and MSCs-derived exosomes to assess the effect of lncRNA H19/miR-152-3p/PTEN axis on the biological activities and inflammation in fibroblasts. Mouse models of DFU were developed by streptozotocin, which were injected with MSCs-derived exosomes overexpressing lncRNA H19. LncRNA H19 in MSCs was transferred through exosomes to fibroblasts, the mechanism of which improved wound healing in DFU, correspond to promoted fibroblast proliferation and migration as well as suppressed apoptosis and inflammation. Wound healing in mice with DFU was facilitated following the injection of MSCs-derived exosomes overexpressing lncRNA H19. Taken together, MSCs-derived exosomal lncRNA H19 prevented the apoptosis and inflammation of fibroblasts by impairing miR-152-3p-mediated PTEN inhibition, leading to stimulated the wound healing process in DFU.

    更新日期:2019-12-17
  • MSTN attenuates cardiac hypertrophy through inhibition of excessive cardiac autophagy by blocking AMPK/mTOR and miR-128/PPARγ/NF-κB signaling pathways
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-14
    Hanping Qi; Jing Ren; Lina Ba; Chao Song; Qianhui Zhang; Yonggang Cao; Pilong Shi; Bowen Fu; Yongsheng Liu; Hongli Sun

    Cardiac hypertrophy, a response of the heart to increased workload, is a major risk factor for heart failure. Myostatin (MSTN) is an inhibitor of myogenesis, regulating the number and size of skeletal myocytes. In recent years, cardiomyocyte autophagy also has been considered to involve in controlling the hypertrophic response. However, less is known about the detailed mechanism of MSTN on cardiac hypertrophy via regulation of cardiomyocyte autophagy. In this study, we found that the deletion of MSTN potentiated abdominal aorta coarctation (AAC) and Ang II–induced pathological cardiac hypertrophy and cardiac autophagy; however, AAC and Ang II–induced cardiac hypertrophic phenotype and cardiac autophagy were dramatically diminished by MSTN in vivo and in vitro. Mechanistically, the anti-hypertrophic and anti-autophagic effects mediated by MSTN in response to pathological stimuli were associated with the direct inactivation of AMPK/mTOR and activation of the PPARγ/NF-κB signaling pathway. Additionally, miR-128 aggravated the progression of cardiac hypertrophy through suppressing its target PPARγ. Furthermore, MSTN downregulated miR-128 expression induced by AAC and Ang II. Taken together, MSTN significantly blunts pathological cardiac hypertrophy and dysfunction, at least in part, by inhibiting excessive cardiac autophagy via blocking AMPK/mTOR and miR-128/PPARγ/NF-κB signaling pathways.

    更新日期:2019-12-17
  • Melatonin ameliorates post-subarachnoid hemorrhage (SAH) cerebral vasospasm by affecting the expression of endothelial nitric oxide synthase (eNOS) and hypoxia-inducible factor 1α (HIF1α) via H19/miR-138/eNOS/NO and H19/miR-675/HIF1α signaling pathways
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-14
    Guoqiang Hou; Hongjin Chen; Yuhua Yin; Yaohua Pan; Xiaohua Zhang; Feng Jia

    Melatonin (MEL) has been demonstrated to exert a protective effect against subarachnoid hemorrhage (SAH), and nitric oxide (NO) has been shown to play an important role in the pathogenesis of vasospasm. This study aims to explore the underlying molecular mechanisms of MEL in the control of vasospasm following SAH. MEL administration attenuates SAH-induced vasospasm and neurobehavioral deficits. Expressions of H19, eNOS and miR-675 are low in the SAH group, while expressions of miR-138 and HIF1α are high in the SAH group. Also, MEL treatment upon SAH rats completely restores the dysregulation of H19, eNOS, miR-675, miR-138 and HIF1α to their normal levels. Moreover, MEL dose-dependently increases the luciferase activity of H19 promoter and hence the expression of H19. Additionally, H19 directly targets miR-675 and miR-138 to increase miR-675 expression and inhibit miR-138 expression. As virtual target genes of miR-675 and miR-138, respectively, HIF1α and eNOS are also regulated by the treatment with MEL. In particular, MEL treatment increases the expression of miR-675 and eNOS level, while decreasing the expression of miR-138 and HIF1α in a dose-dependent manner. Our study found that MEL ameliorates post-SAH vasospasm by regulating the expression of eNOS and HIF1α via H19/miR-138/eNOS/NO and H19/miR-675/HIF1α signaling pathways.

    更新日期:2019-12-17
  • m6A methylation of precursor-miR-320/RUNX2 controls osteogenic potential of bone marrow-derived mesenchymal stem cells
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-12
    Gege Yan; Ye Yuan; Mingyu He; Rui Gong; Hong Lei; Hongbao Zhou; Wenbo Wang; Weijie Du; Tianshuai Ma; Shenzhen Liu; Zihang Xu; Manqi Gao; Meixi Yu; Yu Bian; Ping Pang; Xin Li; Shuting Yu; Fan Yang; Lei Yang
    更新日期:2019-12-13
  • MicroRNA-129 Inhibits Glioma Cells Growth by Targeting CDK4, CDK6 and MDM2
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-12
    Atieh Moradimotlagh; Ehsan Arefian; Rezvan Rezazadeh Valojerdi; Shokoofeh Ghaemi; Fatemeh Jamshidi Adegani; Masoud Soleimani

    Glioblastoma is the most common malignant primary brain tumor among adults and one of the most lethal cancers. It is characterized by the deregulation of signaling pathways involving proliferation, growth, survival, etc. MiRNAs play a role in gene’s regulation by affecting the 3´UTR region of mRNA and affect many cell functions. The present study showed that miR-129 decreased the expression of retinoblastoma and p53 signaling pathways’ genes, including CDK4, CDK6, and MDM2. The Real-time PCR data indicated that expression of CDK4 in U251 and U87 cell lines declined by 69.8% and 47% (p<0.05) respectively, and expression of CDK6 and MDM2 in U251 cells decreased by 55.3% (p<0.0001) and 34.7% (p<0.05) significantly. Luciferase assay confirmed that, overexpression of miR-129 decreased the expression of CDK4 gene by 58.9% (p<0.01), CDK6 by 35.7% (p<0.0001) and MDM2 by 49% (p<0.001). Moreover, cell cycle assay showed a decrease of G2 phase population to 10% and pre-G2 arrest in U87 cells (p<0.05). Besides, wound healing assay indicated that miR-129 overexpression inhibits cell growth of glioblastoma cells. These findings introduced novel targets for miR-129 in glioblastoma cells.

    更新日期:2019-12-13
  • Silencing long non-coding RNA PVT1 upregulates microRNA-145 and confer inhibitory effects on viability, invasion and migration in esophageal carcinoma through knockdown of FSCN1
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-06
    Si-Ning Shen, Ke Li, Ying Liu, Cheng-Liang Yang, Chun-Yu He, Hao-Rang Wang

    Long non-coding RNA (LncRNA) plasmacytoma variant translocation 1 (PVT1) is correlated to various malignant tumors. Consequently, we explored effects of lncRNA PVT1 on esophageal carcinoma (EC) targeting microRNA-145 (miR-145). EC tissues, adjacent normal tissues, and EC-related cell lines were collected and cultured. Expression of lncRNA PVT1, miR-145, FSCN1 and related genes with intervening expression of PVT1 and miR-145 was determined. Bioinformatic website, dual-luciferase reporter assay and RIP were carried to verify target relationship among lncRNA PVT1, FSCN1 and miR-145. Scratch test, Transwell assay, MTT assay, and Flow cytometry were performed for detection of migration, invasion, viability, apoptosis of transfected cells respectively. Finally, tumor formation in nude mice was measured. After database analysis, lncRNA PVT1, miR-145 and FSCN1 were selected for study. LncRNA PVT1 and FSCN1 can bind to miR-145. After overexpressing miR-145 or inhibiting lncRNA PVT1, EC cell viability, migration, and invasion were inhibited, while volume and weight of tumor formation in nude mice decreased. Expression of lncRNA PVT1, FSCN1, Bcl-2, CD147, VEGFR2, MTA1 decreased and expression of miR-145 and Bax increased. Silencing lncRNA PVT1 can upregulate miR-145, which is a tumor suppressor in EC via knockdown of FSCN1. Thus, we might provide a potential theoretical basis for EC treatment.

    更新日期:2019-12-06
  • Mesenchymal stem cells-secreted exosomal H19 promotes trophoblast cell invasion and migration by down-regulating let-7b and up-regulating FOXO1 via AKT signaling pathway in preeclampsia
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-06
    Yang Chen, Haiyan Ding, Min Wei, Wenhui Zha, Shuang Guan, Ning Liu, Yang Li, Yuan Tan, Yan Wang, Fuju Wu

    Exosomes perform important functions for intercellular communication through extracellular signaling pathways, leading to the regulation of important biological processes including cell proliferation but also, systemic dysfunctions such as preeclampsia (PE). However, the inhibitory effects of mesenchymal stem cells (MSCs)-derived exosomes in PE remain largely unknown. Thus, we assessed the possibility that exosomes could transport long noncoding RNA H19 and the correlation between H19 and the apoptosis of trophoblast cells. The expression of microRNA let-7b and forkhead box protein O1 (FOXO1) was characterized in placental tissues of PE patients. Gain- and loss-of-function experiments were performed to examine the roles of FOXO1 and let-7b in trophoblast cells. Interactions between let-7b and H19 as well as between let-7b and FOXO1 were confirmed by a dual luciferase reporter assay, RNA-pull down and, RNA immunoprecipitation. HTR-8/SVNEO cells were co-cultured with exosomes derived from MSCs overexpressing H19, followed by invasion, migration and apoptosis assessments of trophoblast cells. We found that Let-7b was highly expressed and FOXO1 was poorly expressed in placental tissues of PE patients. Furthermore, H19 acts as a competitive endogenous RNA against let-7b, and let-7b directly targeted FOXO1. Moreover, H19 could be transferred to trophoblast cells via MSCs-secreted exosomes. MSCs-derived exosomes overexpressing H19 decreased let-7b, increased FOXO1 and activated the protein kinase B (AKT) signaling pathway, thus increasing invasion and migration, and inhibiting apoptosis of trophoblast cells. These results suggest that MSCs-derived exosomes overexpressing H19 may be a novel direction for therapeutic strategies against PE.

    更新日期:2019-12-06
  • Circular RNA circSLC26A4 accelerates cervical cancer progression via miR-1287-5p/HOXA7 axis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-12-06
    Fei Ji, Rong Du, Tianfeng Chen, Meng Zhang, Yuanfang Zhu, Xin Luo, Yan Ding

    Circular RNAs (circRNAs) are group of non-coding RNAs derived from back-splicing events. Accumulating evidence certificates the critical roles of circRNAs in human tumorigenesis. However, the role and biogenesis of circRNAs in the cervical cancer are still unclear. Here, a novel identified circRNA, circSLC26A4, was found to be up-regulated in cervical cancer tissue and cells. Clinically, the high-expression of circSLC26A4 was related to the poor survival of cervical cancer patients. Functionally, cellular experiments indicated that circSLC26A4 knockdown repressed the proliferation, invasion and tumor growth in vitro and in vivo. Furthermore, circSLC26A4 acted as the sponge of miR-1287-5p, moreover miR-1287-5p targeted the 3’-UTR of HOXA7 mRNA. Mechanistically, RNA binding protein (RBP) Quaking (QKI) was identified to interact with the QKI response elements (QREs) in SLC26A4 gene introns, thereby promoting circSLC26A4 biogenesis. In conclusion, these findings demonstrate that circSLC26A4 facilitates the cervical cancer progression through QKI/circSLC26A4/miR-1287-5p/HOXA7 axis, which might bring novel therapeutic strategies for cervical cancer.

    更新日期:2019-12-06
  • MiRNA-129/FBW7/NFkB, a novel regulatory pathway in inflammatory bowel disease
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-30
    Qinghui Meng, Weihua Wu, Tiemin Pei, Junlin Xue, Peng Xiao, Liang Sun, Long Li, Desen Liang

    F-box and WD repeat domain containing protein 7 (FBW7) has been documented to be implicated in NFκB signaling and inflammation, but its role in the pathogenesis of inflammatory bowel disease (IBD) remains unknown. FBW7 was increased both in colon tissues from IBD patients and trinitrobenzene sulphonic acid (TNBS)-induced colitis mice. Immunoprecipitation assay identified that FBW7 as a novel IκBα-binding partner. FBW7 upregulation promoted IκBα ubiquitin-dependent degradation, NFκB activation and subsequent intestinal inflammation in intestinal epithelial cells, whereas inhibition of FBW7 produced the opposite effects. Computational analysis revealed that miR-129 directly targets at 3’UTR of FBW7. MiR-129 suppressed proteasome pathway-mediated the degradation of IκBα by negatively regulating FBW7. The in vivo study demonstrated that upregulation of miR-129 ameliorated intestinal inflammation in TNBS-induced colitis mice through inhibition NFκB signaling pathway. In conclusion, FBW7 is a novel E3 ubiquitin ligase for IκBα, thereby leads to NFκB activation and inflammation. MiR-129 negatively regulates FBW7 expression, resulting in secondary inhibition of NFκB pathway and amelioration of intestinal inflammation. Our findings provide a new insight into the development of therapeutic strategies for the treatment of IBD.

    更新日期:2019-11-30
  • The LINC01714 enhances Gemcitabine sensitivity by modulating FOXO3 phosphorylation in cholangiocarcinoma
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-30
    Sheng Shen, Jiwen Wang, Bohao Zheng, Ying Tao, Min Li, Yueqi Wang, Xiaoling Ni, Tao Suo, Houbao Liu, Han Liu, Jiwei Zhang

    Long noncoding RNAs (lncRNAs) have been shown to play crucial roles in human cancers. However, the underlying biological functions and mechanisms of lncRNAs in cholangiocarcinoma (CCA) remain largely unknown. We aimed to characterize transcriptional landscape of lncRNAs in CCA and identify lncRNAs that were able to serve as prognosis markers and therapeutic targets for CCA. Here, we investigated the transcriptional landscape and dysregulation of lncRNAs in CCA. LINC01714 was found to be recurrently down-regulated in CCA tumor samples. Our results revealed that decreased LINC01714 expression was associated with poor survival of CCA patients. Our observations revealed that LINC01714 suppressed the proliferation, migration and invasion abilities of CCA cells both in vitro and in vivo. Furthermore, we found that LINC01714 physically interacted with Forkhead Box O3 (FOXO3) and increased FOXO3 protein level. In addition, LINC01714 could decrease the phosphorylation level of FOXO3. Interestingly, LINC01714 was able to enhance the sensitivity of Gemcitabine on CCA tumor cells through modulating phosphorylated FOXO3-Ser318. Our study revealed LINC01714 as a promising prognostic indictor for patients with CCA, provided insights into the molecular pathogenesis of CCA and also showed that LINC01714 is a potential therapeutic combination for Gemcitabine in CCA treatment.

    更新日期:2019-11-30
  • Plk2 regulated by miR-128 induces ischemia reperfusion injury in cardiac cells
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-30
    Duo Zhao, Edward Shun, Fengjuling, Qing Liu, Ayesha Warsi, Bowen Wang, Qinfeng Zhou, Cuilin Zhu, Hao Zheng, Kexiang Liu, Xiufen Zheng

    Ischemia-reperfusion (I/R) injury occurs during cardiac surgery and is the major factor leading to heart dysfunction and heart failure. Our previous study showed that gene and microRNA expression profiles are altered in heart grafts with extended ischemia reperfusion injury. In this study, we, for the first time, demonstrated that I/R injury upregulates the expression of Plk2 but decreases miR-128 expression in heart cells both in vitro and in vivo. Silencing Plk2 using siRNA protects cells from Antimycin A induced cell apoptosis/death. Silencing Plk2 also decreases phosphorylated p65 expression but increases Angiopoietin 1 expression. Plk2 is negatively regulated by miR-128. miR-128 exerts a similar protective effect on cell apoptosis/death to Plk2 siRNA in heart cells in response to I/R stress. Methylation inhibitor 5-Azacytidine (5-AZ) increases the expression of miR-128 and subsequently reduces Plk2 expression and cell apoptosis. In conclusion, this study demonstrated that Plk2 regulated by miR-128 induces cell apoptosis/death in response I/R stress through activation of the NFkB signal pathway. miR-128 and Plk2 are new targets for preventing cardiac I/R injury or oxidative stress mediated injury.

    更新日期:2019-11-30
  • LINC01413/hnRNP-K/ZEB1 axis accelerates cell proliferation and epithelial-mesenchymal transition in colorectal cancer via inducing YAP1/TAZ1 translocation
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-29
    Ling Ji, Xiang Li, Zhenhua Zhou, Zhihai Zheng, Li Jin, Feizhao Jiang

    Long non-coding RNAs (lncRNAs) are crucial molecules in tumorigenesis and tumor growth in various human cancers, including colorectal cancer (CRC). Studies have revealed that lncRNAs can regulate cellular processes in cancers by interacting with proteins, for example RNA binding proteins (RBPs). Herein, we recognized a novel lncRNA called LINC01413 which is up-regulated in CRC tissues through lncRNAs microarray. Subsequently, it is confirmed that elevated level of LINC01413 expression in CRC tissues and was strongly correlated to clinicopathological features, such as tumor size, tumor stage, lymph node metastasis, and distant metastasis, and its association with poor overall survival was also revealed. Additionally, LINC01413 facilitates cell proliferation, migration, invasion, and EMT in vitro. Also, silenced LINC01413 restrains tumor growth in vivo. Moreover, LINC01413 binds with hnRNP-K and induces YAP1/TAZ1 nuclear translocation to regulate the expression of ZEB1 in CRC cells. Together, this research suggested LINC01413 as a positive regulator in CRC progression through LINC01413/hnRNP-K/TAZ1/YAP1/ZEB1 axis, broadening a new view on CRC treatment.

    更新日期:2019-11-29
  • LncRNA UCA1 promotes gefitinib resistance as a ceRNA to target FOSL2 by sponging miR-143 in non-small cell lung cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-29
    Xiliu Chen, Zhongliang Wang, Fan Tong, Xiaorong Dong, Gang Wu, Ruiguang Zhang

    Exosomes could mediate cell-cell crosstalk in cancer progression by transferring long noncoding RNAs (lncRNAs). The aim of this study is to explore the roles of the exosomal lncRNA urothelial carcinoma-associated (UCA1) on gefitinib resistance in non-small-cell lung cancer (NSCLC). First, we detected the expression of UCA1 in gefitinib resistant and gefitinib sensitive NSCLC by qRT-PCR; the expression occurred in tissues, cell lines and exosomes. Cell phenotypes and animal experiments were performed to determine the effect of UCA1 and exosomal UCA1. Furthermore, bioinformatics online programs and luciferase reporter assay were used to validate the association of UCA1 and miR-143 in NSCLC cells. We observed that UCA1 was increased in both gefitinib resistant NSCLC cells and their secreted exosomes. In vitro and in vivo experiments demonstrated that UCA1 knockdown impaired cell proliferation and promoted the gefitinib-induced cell apoptosis. Then we demonstrated that repressed UCA1 promoted the miR-143 expression and miR-143 could bind to the predicted binding site of UCA1. We then dissected the effect of miR-143 on gefitinib resistance in NSCLC and proved the suppressive role of miR-143. Furthermore, we found that miR-143 displayed its role via modulating the FOSL2 expression. In summary, our findings indicate that exosomal UCA1 may serve as a promising therapeutic target for the treatment of EGFR+ NSCLC patients.

    更新日期:2019-11-29
  • H19 increases IL-17A/IL-23 releases via regulating VDR by interacting with miR675-5p/miR22-5p in ankylosing spondylitis
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-29
    Xu Zhang, Shuang Ji, Guoqi Cai, Zhipeng Pan, Renfang Han, Yaping Yuan, Shanshan Xu, Jiajia Yang, Xingxing Hu, Mengya Chen, Meng Wu, Yubo Ma, Jixiang Deng, Xing Gao, Shiyang Guan, Shengqian Xu, Zongwen Shuai, Laura Laslett, Faming Pan

    Long non-coding RNA (LncRNA) H19 is associated with inflammatory diseases but the molecular mechanism of H19 in the inflammatory process of ankylosing spondylitis (AS) is unclear. Here we investigated the role of H19 and its downstream molecules in the inflammation of AS by microarrays analysis, qRT-PCR, western blot, and dual-luciferase reporter assay. H19 siRNA (Si-H19) and adenovirus (AD-H19) were used to decrease and increase H19 expression, respectively. 42 annotated lncRNAs were identified and H19 was overexpressed. H19, vitamin D receptor (VDR), and transforming growth factor (TGF)-β can bind to microRNA (miR)22-5p and miR675-5p. Si-H19 significantly down-regulated miR22-5p and up-regulated miR675-5p expression; Si-H19 decreased the protein and mRNA expression of VDR and decreased the cytokine and mRNA levels of interleukin (IL)-17A and IL-23. These results were verified by AD-H19. In addition, miR22-5p and miR675-5p inhibitors increased the protein and mRNA expression of VDR and increased the cytokine and mRNA levels of IL-17A and IL-23. These results were also confirmed by miRNA mimics. Furthermore, H19 directly interfered with miR22-5p and miR675-5p expression, whereas the two miRNAs directly inhibited VDR expression. Overall, the H19-miR22-5p/miR675-5p-VDR-IL-17A/IL-23 signaling pathways have important roles in the pathogenesis of AS.

    更新日期:2019-11-29
  • CircRNAs and exosomes: a mysterious frontier for human cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-29
    Xuefei Shi, Bin Wang, Xuereng Feng, Yongcan Xu, Kaihua Lu, Ming Sun

    Exosomes are nano-size membrane-bound vesicles and contain active substances (DNA, noncoding RNA, protein), which provide a novel method of transferring effector messages between cells. Circular RNAs (circRNAs), a kind of ncRNAs, have attracted increasing attention over the last decade given advances in whole genome and transcriptome sequencing technologies. It has become increasingly clear that circRNAs regulate gene expression through various actions and play diverse roles in many fields of human cancer biology. Notably, several studies reported that circRNAs are enriched in exosomes and that exosomal circRNAs play an important role in cancer biology. Exosomal circRNAs can be taken up by neighboring or distant cells and affect many aspects of physiological and pathological conditions of the recipient cells, potentially promoting cell communication and tumor metastasis. Here, we briefly review the molecular mechanisms of circRNAs and recent findings regarding exosomal circRNAs, and highlight the specific roles of exosomal circRNAs in human cancer.

    更新日期:2019-11-29
  • microRNA-181d-5p-containing exosomes derived from cancer-associated fibroblasts promote epithelial-mesenchymal transition by regulating CDX2/HOXA5 in breast cancer
    Mol. Ther. Nucl. Acids (IF 5.919) Pub Date : 2019-11-29
    Hongbin Wang, Hong Wei, Jingsong Wang, Lin Li, Anyue Chen, Zhigao Li

    Recently, novel mechanisms underlying the pro-tumorigenic effects of cancer-associated fibroblasts (CAFs) have been identified in several cancers, including breast cancer. CAFs can secrete exosomes which are loaded with proteins, lipids and RNAs to affect tumor microenvironment. Herein we identify CAFs-derived exosomes that can transfer miR-181d-5p to enhance the aggressiveness of breast cancer. Cancerous tissues and matched paracancerous tissues were surgically resected from 122 patents with breast cancer. ChIP and dual luciferase reporter assays were employed to identify interaction between HOXA5 and CDX2 as well as between CDX2 and miR-181d-5p respectively. Human breast cancer MCF-7 cells were co-cultured with CAFs-derived exosomes. EdU assay, TUNEL staining, Transwell invasion assays, and scratch tests were carried out to evaluate MCF-7 cell functions. Nude mice bearing xenografted MCF-7 cells were injected with CAFs-derived exosomes and the tumor formation was evaluated. HOXA5 expressed at a poor level in breast cancer tissues, and its overexpression retarded MCF-7 cell proliferation, invasion, migration and epithelial-mesenchymal transition (EMT) and facilitated its apoptosis in vitro. miR-181d-5p targets CDX2, a transcription factor binding to HOXA5 promoter. Coculturue of CAFs and MCF-7 cells showed CAFs prolonged proliferation and antagonized apoptosis of MCF-7 cells via release of exosomes. Coculturue of MCF-7 cells and exosomes derived from CAFs identified miR-181d-5p as a mediator of the exosomal effects on MCF-7 cells, in part via down-regulation of CDX2 and HOXA5. CAFs-derived exosomes containing miR-181d-5p decreased the tumor growth of nude mice bearing xenografted MCF-7 cells. In conclusion, exosomal miR-181d-5p plays a key role in CAFs-mediated effects on tumor environment in breast cancer, likely via CDX2 and HOXA5.

    更新日期:2019-11-29
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