Abstract
Glioma is a common malignant brain neoplasm. The role and mechanism of circular RNA 0,007,534 (circ_0007534) in glioma progression were investigated in this study. The expression of circ_0007534, microRNA-22-3p (miR-22-3p) and prospero homeobox protein 1 (PROX1) messenger RNA (mRNA) was determined by quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation, migration and invasion abilities were analyzed by colony formation assay, transwell migration assay and transwell invasion assay. Cell apoptosis was assessed through measuring the activity of Caspase-3 using the Caspase-3 kit and the apoptosis rate using flow cytometry. Dual-luciferase reporter assay was used to confirm the target interaction between miR-22-3p and circ_0007534 or PROX1. The protein level of PROX1 was examined by Western blot assay. Animal studies were conducted to analyze the influence of circ_0007534 interference on xenograft tumor growth in vivo. Circ_0007534 was highly expressed in glioma tissues and cell lines relative to that in normal tissues and NHA cell line. Circ_0007534 knockdown suppressed the proliferation and motility while induced the apoptosis of glioma cells. Circ_0007534 negatively regulated miR-22-3p level through targeting it in glioma cells. Circ_0007534 interference-induced influences in glioma cells were partly overturned by the silencing of miR-22-3p. PROX1 was a target of miR-22-3p, and circ_0007534 interference-mediated effects in glioma cells were largely diminished by the overexpression of PROX1. Circ_0007534 interference restrained glioma development in vivo. Circ_0007534 aggravated glioma progression through elevating PROX1 expression via targeting miR-22-3p, which provided new targets for the diagnosis and treatment of glioma.
Similar content being viewed by others
Data Availability
The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.
References
Anastasiadou E, Jacob LS, Slack FJ (2018) Non-coding RNA networks in cancer. Nat Rev Cancer 18(1):5–18
Cui P, Su J, Li Q, Xu G, Zhu N (2019) LncRNA RHPN1-AS1 targeting miR-625/REG3A promotes cell proliferation and invasion of glioma cells. Onco Targets Ther 12:7911–7921
Duan X, Liu D, Wang Y, Chen Z (2018) Circular RNA hsa_circ_0074362 promotes glioma cell proliferation, migration, and invasion by attenuating the inhibition of miR-1236-3p on HOXB7 expression. DNA Cell Biol 37(11):917–924
Elsir T, Eriksson A, Orrego A, Lindström MS, Nistér M (2010) Expression of PROX1 Is a common feature of high-grade malignant astrocytic gliomas. J Neuropathol Exp Neurol 69(2):129–138
Elsir T, Qu M, Berntsson SG, Orrego A, Olofsson T, Lindström MS et al (2011) PROX1 is a predictor of survival for gliomas WHO grade II. Br J Cancer 104(11):1747–1754
Fabian MR, Sonenberg N, Filipowicz W (2010) Regulation of mRNA translation and stability by microRNAs. Annu Rev Biochem 79:351–379
Fan L, Wu Y, Wang J, He J, Han X (2019) Sevoflurane inhibits the migration and invasion of colorectal cancer cells through regulating ERK/MMP-9 pathway by up-regulating miR-203. Eur J Pharmacol 850:43–52
Hansen TB, Jensen TI, Clausen BH, Bramsen JB, Finsen B, Damgaard CK et al (2013) Natural RNA circles function as efficient microRNA sponges. Nature 495(7441):384–388
Hao L, Rong W, Bai L, Cui H, Zhang S, Li Y et al (2019) Upregulated circular RNA circ_0007534 indicates an unfavorable prognosis in pancreatic ductal adenocarcinoma and regulates cell proliferation, apoptosis, and invasion by sponging miR-625 and miR-892b. J Cell Biochem 120(3):3780–3789
Hu T, Wang F, Han G (2020) LncRNA PSMB8-AS1 acts as ceRNA of miR-22–3p to regulate DDIT4 expression in glioblastoma. Neurosci Lett 728:134896
Jiang W, Han X, Wang J, Wang L, Xu Z, Wei Q et al (2019) miR-22 enhances the radiosensitivity of small-cell lung cancer by targeting the WRNIP1. J Cell Biochem 120(10):17650–17661
Kim H, Cruz M, Bourdeau A, Dumont DJ (2013) Cell-cell interactions influence vascular reprogramming by Prox1 during embryonic development. PLoS ONE 8(1):e52197
Kristensen LS, Hansen TB, Venø MT, Kjems J (2018) Circular RNAs in cancer: opportunities and challenges in the field. Oncogene 37(5):555–565
Lei B, Huang Y, Zhou Z, Zhao Y, Thapa AJ, Li W et al (2019) Circular RNA hsa_circ_0076248 promotes oncogenesis of glioma by sponging miR-181a to modulate SIRT1 expression. J Cell Biochem 120(4):6698–6708
Li B, Li X (2018) Overexpression of hsa_circ_0007534 predicts unfavorable prognosis for osteosarcoma and regulates cell growth and apoptosis by affecting AKT/GSK-3β signaling pathway. Biomed Pharmacother 107:860–866
Li GF, Li L, Yao ZQ, Zhuang SJ (2018) Hsa_circ_0007534/miR-761/ZIC5 regulatory loop modulates the proliferation and migration of glioma cells. Biochem Biophys Res Commun 499(4):765–771
Liang M, Huang G, Liu Z, Wang Q, Yu Z, Liu Z et al (2019) Elevated levels of hsa_circ_006100 in gastric cancer promote cell growth and metastasis via miR-195/GPRC5A signalling. Cell Prolif 52(5):e12661
Liu DK, Wang J, Guo Y, Sun ZX, Wang GH (2019) Identification of differentially expressed genes and fusion genes associated with malignant progression of spinal cord gliomas by transcriptome analysis. Sci Rep 9(1):13583
Liu M, Xu A, Yuan X, Zhang Q, Fang T, Wang W et al (2015a) Downregulation of microRNA-409-3p promotes aggressiveness and metastasis in colorectal cancer: an indication for personalized medicine. J Transl Med 13:195
Liu Y, Ye X, Zhang JB, Ouyang H, Shen Z, Wu Y et al (2015b) PROX1 promotes hepatocellular carcinoma proliferation and sorafenib resistance by enhancing β-catenin expression and nuclear translocation. Oncogene 34(44):5524–5535
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402–408
Lu Y, Deng X, Xiao G, Zheng X, Ma L, Huang W (2019) circ_0001730 promotes proliferation and invasion via the miR-326/Wnt7B axis in glioma cells. Epigenomics 11(11):1335–1352
Meng S, Zhou H, Feng Z, Xu Z, Tang Y, Li P et al (2017) CircRNA: functions and properties of a novel potential biomarker for cancer. Mol Cancer 16(1):94
Milano MT, Johnson MD, Sul J, Mohile NA, Korones DN, Okunieff P et al (2010) Primary spinal cord glioma: a surveillance, epidemiology, and end results database study. J Neurooncol 98(1):83–92
Panda AC (2018) Circular RNAs Act as miRNA sponges. Adv Exp Med Biol 1087:67–79
Qi X, Zhang DH, Wu N, Xiao JH, Wang X, Ma W (2015) ceRNA in cancer: possible functions and clinical implications. J Med Genet 52(10):710–718
Qi Y, Zhang B, Wang J, Yao M (2018) Upregulation of circular RNA hsa_circ_0007534 predicts unfavorable prognosis for NSCLC and exerts oncogenic properties in vitro and in vivo. Gene 676:79–85
Qian L, Guan J, Wu Y, Wang Q (2019) Upregulated circular RNA circ_0074027 promotes glioblastoma cell growth and invasion by regulating miR-518a-5p/IL17RD signaling pathway. Biochem Biophys Res Commun 510(4):515–519
Rong X, Gao W, Yang X, Guo J (2019) Downregulation of hsa_circ_0007534 restricts the proliferation and invasion of cervical cancer through regulating miR-498/BMI-1 signaling. Life Sci 235:116785
See SJ, Gilbert MR (2004) Anaplastic astrocytoma: diagnosis, prognosis, and management. Semin Oncol 31(5):618–634
Sha QK, Chen L, Xi JZ, Song H (2019) Long non-coding RNA LINC00858 promotes cells proliferation, migration and invasion by acting as a ceRNA of miR-22-3p in colorectal cancer. Artif Cells Nanomed Biotechnol 47(1):1057–1066
Siegel RL, Miller KD, Jemal A (2019) Cancer statistics. CA Cancer J Clin 69(1):7–34
Skog M, Bono P, Lundin M, Lundin J, Louhimo J, Linder N et al (2011) Expression and prognostic value of transcription factor PROX1 in colorectal cancer. Br J Cancer 105(9):1346–1351
Song L, Xiao Y (2018) Downregulation of hsa_circ_0007534 suppresses breast cancer cell proliferation and invasion by targeting miR-593/MUC19 signal pathway. Biochem Biophys Res Commun 503(4):2603–2610
Sosa-Pineda B, Wigle JT, Oliver G (2000) Hepatocyte migration during liver development requires Prox1. Nat Genet 25(3):254–255
Sun FB, Lin Y, Li SJ, Gao J, Han B, Zhang CS (2018) MiR-210 knockdown promotes the development of pancreatic cancer via upregulating E2F3 expression. Eur Rev Med Pharmacol Sci 22(24):8640–8648
Ueta K, Otowa Y, Kakeji Y, Hirashima M (2018) PROX1 is associated with cancer progression and prognosis in gastric cancer. Anticancer Res 38(11):6139–6145
van Tellingen O, Yetkin-Arik B, de Gooijer MC, Wesseling P, Wurdinger T, de Vries HE (2015) Overcoming the blood-brain tumor barrier for effective glioblastoma treatment. Drug Resist Updat 19:1–12
Wei Y, Lu C, Zhou P, Zhao L, Lyu X, Yin J et al (2021) EIF4A3-induced circular RNA ASAP1 promotes tumorigenesis and temozolomide resistance of glioblastoma via NRAS/MEK1/ERK1-2 signaling. Neuro Oncol 23(4):611–624
Xu X, Wan X, Wei X (2017) PROX1 promotes human glioblastoma cell proliferation and invasion via activation of the nuclear factor-κB signaling pathway. Mol Med Rep 15(2):963–968
Zhang B, Ji S, Ma F, Ma Q, Lu X, Chen X (2016) miR-489 acts as a tumor suppressor in human gastric cancer by targeting PROX1. Am J Cancer Res 6(9):2021–2030
Zhang F, Ruan X, Ma J, Liu X, Zheng J, Liu Y et al (2020) DGCR8/ZFAT-AS1 promotes CDX2 transcription in a PRC2 complex-dependent manner to facilitate the malignant biological behavior of glioma cells. Mol Ther 28(2):613–630
Zhang HD, Jiang LH, Sun DW, Hou JC, Ji ZL (2018a) CircRNA: a novel type of biomarker for cancer. Breast Cancer 25(1):1–7
Zhang R, Wei RL, Du W, Zhang LW, Du T, Geng YD et al (2019) Long noncoding RNA ENST00000413528 sponges microRNA-593-5p to modulate human glioma growth via polo-like kinase 1. CNS Neurosci Ther 25(8):842–854
Zhang R, Xu J, Zhao J, Wang X (2018b) Silencing of hsa_circ_0007534 suppresses proliferation and induces apoptosis in colorectal cancer cells. Eur Rev Med Pharmacol Sci 22(1):118–126
Zhang W, Yang S, Liu Y, Wang Y, Lin T, Li Y et al (2018c) Hsa_circ_0007534 as a blood-based marker for the diagnosis of colorectal cancer and its prognostic value. Int J Clin Exp Pathol 11(3):1399–1406
Zinovieva RD, Duncan MK, Johnson TR, Torres R, Polymeropoulos MH, Tomarev SI (1996) Structure and chromosomal localization of the human homeobox gene Prox 1. Genomics 35(3):517–522
Funding
The project was funded by Shenzhen Sanming Project (SZSM201806067).
Author information
Authors and Affiliations
Contributions
RM designed and supervised the study. YZ conducted the experiments and drafted the manuscript. LL conducted the experiments and supervised the study. ZZ collected and analyzed the data. YW contributed the methodology and analyzed the data. YC edited the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical Approval
This study was approved by the Ethics Review Committee of the Second Affiliated Hospital of Shenzhen University and was carried out according to the guidelines of Declaration of Helsinki.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
10571_2021_1150_MOESM1_ESM.tif
Supplementary file1 Supplementary Figure 1. Circ_0007534 silencing has almost no effect on the apoptosis of human astrocytes NHA. Flow cytometry was performed to analyze the apoptosis rate in NHA cells with the silencing of circ_0007534 or not (TIF 771 kb)
10571_2021_1150_MOESM2_ESM.tif
Supplementary file2 Supplementary Figure 2. The candidate miRNA targets of circ_0007534. (A and B) The expression of miR-1236, miR-593, miR-22-3p, miR-625, and miR-502-5p was determined by qRT-PCR in LN229 and U251 cells transfected with si-NC or si-circ_0007534. *P<0.05, **P<0.01, ***P<0.001 (TIF 258 kb)
10571_2021_1150_MOESM3_ESM.tif
Supplementary file3 Supplementary Figure 3. The candidate mRNA targets of miR-22-3p. (A and B) qRT-PCR was conducted to detect the levels of GRM5, IL17RD, PROX1, and CDX2 in LN229 and U251 cells transfected with anti-NC or anti-miR-22-3p. *P<0.05, **P<0.01, ***P<0.001 (TIF 290 kb)
10571_2021_1150_MOESM4_ESM.tif
Supplementary file4 Supplementary Figure 4. PROX1 knockdown triggers the apoptosis of glioma cells. (A) The apoptosis of glioma cells transfected with si-NC or si-PROX1 was analyzed via flow cytometry. (B) Western blot assay was conducted to measure the expression of proteins associated with cell proliferation (Cyclin D1), motility (MMP2 and MMP9), and apoptosis (Bax). ***P<0.001 (TIF 809 kb)
Rights and permissions
About this article
Cite this article
Zheng, Y., Wang, Y., Mai, R. et al. Circ_0007534 Silencing Inhibits the Proliferation, Migration and Invasion and Induces the Apoptosis of Glioma Cells Partly Through Down-Regulating PROX1 Via Elevating miR-22-3p Level. Cell Mol Neurobiol 42, 2819–2832 (2022). https://doi.org/10.1007/s10571-021-01150-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-021-01150-y