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Circ_0004018 suppresses cell proliferation and migration in hepatocellular carcinoma via miR-1197/PTEN/PI3K/AKT signaling pathway
Cell Cycle ( IF 3.4 ) Pub Date : 2021-09-27 , DOI: 10.1080/15384101.2021.1962633
He Wang 1 , Qiao Zhang 2 , Wenyu Cui 2 , Wenlan Li 2 , Jimei Zhang 3
Affiliation  

ABSTRACT

Hepatocellular carcinoma (HCC) is a common type of primary liver cancer. Circular RNAs (circRNAs) have been demonstrated to be a crucial player in multiple cancers. However, a large number of circRNAs remain to be explored. Our study focused on investigating hsa_circ_0004018 in HCC. Firstly, we conducted quantitative reverse transcription PCR (RT-qPCR) to find that circ_0004018 was down-regulated in HCC cells. Western blot analysis was performed to detect the protein levels of phosphatase and tensin homologue (PTEN) and related factors of PI3K/AKT signaling pathway. From the results of functional assays, we found that overexpression of circ_0004018 significantly inhibited the proliferative and migratory capacities of HCC cells. The regulatory mechanism of circ_0004018 in HCC was determined by RNA immunoprecipitation (RIP), RNA pull-down, and luciferase reporter assays, thereby we knew that circ_0004018 regulated PTEN by sequestering microRNA-1197 (miR-1197) to modulate PI3K/AKT signaling pathway. Finally, rescue assays verified that circ_0004018 participated in modulation of cell proliferation and migration in HCC via sponging miR-1197 and regulating PTEN. In conclusion, circ_0004018 suppresses the proliferation and migration of HCC cells via sponging miR-1197 to inactivate the PTEN/PI3K/AKT signaling pathway.

Abbreviations: HCC: Hepatocellular carcinoma; circRNAs: Circular RNAs; PTEN: Phosphatase and tensin homologue; miR-1197: microRNA-1197; ceRNA: competitive endogenous RNA; ATCC: American Type Culture Collection; EMEM: Eagle’s Minimum Essential Medium; RT-qPCR: Quantitative real-time PCR; EdU: 5-ethynyl-20-deoxyuridine; FISH: Fluorescent in situ hybridization; RIP: RNA immunoprecipitation; 3ʹ-UTR: 3ʹ-untranslated region; Wt: wild-type; Mut; mutant type; gDNA: genomic DNA; Act D: Actinomycin D; PI3K: phosphatidylinositol-3-kinase; AKT: protein kinase; lncRNAs: long non-coding RNAs



中文翻译:

Circ_0004018 通过 miR-1197/PTEN/PI3K/AKT 信号通路抑制肝癌细胞增殖和迁移

摘要

肝细胞癌(HCC)是一种常见的原发性肝癌。环状 RNA (circRNA) 已被证明是多种癌症的关键参与者。然而,仍有大量的 circRNA 有待探索。我们的研究重点是调查 HCC 中的 hsa_circ_0004018。首先,我们进行定量逆转录PCR(RT-qPCR)发现circ_0004018在HCC细胞中下调。采用蛋白质印迹法检测磷酸酶和张力蛋白同源物(PTEN)蛋白水平及PI3K/AKT信号通路相关因子。从功能测定的结果中,我们发现circ_0004018的过表达显着抑制了HCC细胞的增殖和迁移能力。circ_0004018 在 HCC 中的调控机制通过 RNA 免疫沉淀 (RIP)、RNA pull-down、和荧光素酶报告基因检测,因此我们知道 circ_0004018 通过隔离 microRNA-1197 (miR-1197) 来调节 PTEN,从而调节 PI3K/AKT 信号通路。最后,救援试验证实 circ_0004018 通过海绵化 miR-1197 和调节 PTEN 参与调节 HCC 中的细胞增殖和迁移。总之,circ_0004018 通过海绵化 miR-1197 使 PTEN/PI3K/AKT 信号通路失活,抑制 HCC 细胞的增殖和迁移。

缩写: HCC:肝细胞癌;circRNAs:环状RNA;PTEN:磷酸酶和张力蛋白同源物;miR-1197:微小RNA-1197;ceRNA:竞争性内源性RNA;ATCC:美国典型培养物保藏中心;EMEM:Eagle 的最低基本培养基;RT-qPCR:定量实时 PCR;EdU:5-乙炔基-20-脱氧尿苷;FISH:荧光原位杂交;RIP:RNA免疫沉淀;3ʹ-UTR:3ʹ-非翻译区;Wt:野生型;穆特; 突变型;gDNA:基因组DNA;D法:放线菌素D;PI3K:磷脂酰肌醇-3-激酶;AKT:蛋白激酶;lncRNA:长链非编码 RNA

更新日期:2021-11-03
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