Effects of miR-1291 on the cell cycle and proliferation of renal cell carcinoma by regulating the expression of Zinc finger protein 8 gene

doi:10.3760/cma.j.issn.1673-422X.2018.03.001

Abstract

Abstract: Objective To investigate the effects of microRNA-1291 (miR-1291) on the expression of Zinc finger protein 8 (PHF8) gene in renal cell carcinoma and its effect on cell cycle and proliferation of renal cell carcinoma. MethodsRealtime fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR1291 in renal cell carcinoma cell lines OS-RC-2, ACHN, A498, 786-O and human proximal tubular epithelial cells HK-2. miR-1291 (miR-1291 group) and miR-NC (miR-NC group) were transfected into the renal cell lines with the lowest expression of miR1291. qRTPCR was used to detect the expression of miR-1291 and PHF8 mRNA in the transfected cells. The expression levels of PHF8, Cyclindependent kinase 6 (CDK6) and Cyclin D1 were detected by Western blotting. The effect of miR-1291 on the transcriptional activity of PHF8 was detected by double luciferase reporter gene system. Flow cytometry was used to detect cell cycle distribution. Methyl thiazolyl tetrazolium (MTT) assay and colony formation assay were used to detect cell viability and proliferation. ResultsThe expressions of miR1291 in renal carcinoma cell lines OS-RC-2, ACHN, A498, 786-O and human proximal renal tubular epithelial cell HK2 were 0.64±0.17, 0.60±0.15, 0.29±0.08, 0.63±0.08 and 1.01±0.17 respectively, with a significant difference (F=13.790, P<0.001). Compared with renal carcinoma cell lines OS-RC-2, ACHN and 786-O, the expression level of miR-1291 in A498 cell line was the lowest (P=0.002, P=0.006, P=0.003). The expression levels of miR1291 in A498 cell lines of miRNC group and miR1291 group were 1.00±0.03 and 775.25±329.91 respectively, with a significant difference (t=4.694, P=0.003); and the expression levels of PHF8 mRNA were 1.00±0.11 and 0.57±0.18 respectively, with a significant difference (t=4.122, P=0.006). The results of Western blotting were consistent with the results of qRTPCR, and the expressions of CDK6 and Cyclin D1 were significantly decreased. The double luciferase reporter gene showed that miR1291 could directly inhibit the activity of luciferase in the 3′ untranslated region of target gene PHF8. Compared with miR-NC group, the proportion of renal carcinoma cells in S phase (23.40±4.29 vs. 32.19±2.64; t=3.491, P=0.013) and G2-M phase (14.38±4.05 vs. 25.59±6.01; t=3.095, P=0.021) decreased; and the proportion of cells in G0-G1 phase increased (62.22±7.56 vs. 42.22±5.23, t=4.351, P=0.005). MTT assay showed that the cell viability of miR1291 was significantly decreased. Colony formation experiments showed that the numbers of colonies formed by A498 cells in miR-NC group and miR1291 group were 246.64±39.94 and 87.34±21.93 respectively, with a significant difference (t=6.993, P<0.001). ConclusionThe expression of miR1291 is significantly decreased in renal cancer cell lines. miR1291 can significantly inhibit the proliferation of renal cell carcinoma cells by targeting interfering PHF8 gene expression, which may contribute to the development of new renal cancer target.

Key words: Kidney neoplasms, MicroRNAs, Cell cycle, Cell proliferation, Zinc finger protein 8

Ye Zhihua, Huang Geng, Fu Jinlun, Gui Dingwen. Effects of miR-1291 on the cell cycle and proliferation of renal cell carcinoma by regulating the expression of Zinc finger protein 8 gene[J]. Journal of International Oncology, 2018, 45(3): 129-133.

References

［1］ Shinohara N, Abe T. Prognostic factors and risk classifications for patients with metastatic renal cell carcinoma［J］. Int J Urol, 2015, 22(10): 888-897. DOI: 10.1111/iju.12858. ［2］ Cutruzzula P, Cahn D, Kivlin D, et al. A review of translocation T(6;11) renal cell carcinoma tumors in the adult patient［J］. Curr Urol, 2017, 10(2): 69-71. DOI: 10.1159/000447154. ［3］ Kanwal R, Plaga AR, Liu X, et al. MicroRNAs in prostate cancer: functional role as biomarkers［J］. Cancer Lett, 2017, 407: 9-20. DOI: 10.1016/j.canlet.2017.08.011. ［4］ Tu MJ, Pan YZ, Qiu JX, et al. MicroRNA-1291 targets the FOXA2AGR2 pathway to suppress pancreatic cancer cell proliferation and tumorigenesis［J］. Oncotarget, 2016, 7(29): 45547-45561. DOI: 10.18632/oncotarget.9999. ［5］ Nakamura K, Sawada K, Yoshimura A, et al. Clinical relevance of circulating cellfree microRNAs in ovarian cancer［J］. Mol Cancer, 2016, 15(1): 48. DOI: 10.1186/s1294301605360. ［6］ Petros FG, Wallis CJD. The utility of microRNAs as biomarkers in predicting progression and survival in patients with clearcell renal cell carcinoma［J］. BJU Int, 2017, 120(3): 305307. DOI: 10.1111/bju.13931. ［7］ Bi HC, Pan YZ, Qiu JX, et al. Nmethylnicotinamide and nicotinamide Nmethyltransferase are associated with microRNA1291altered pancreatic carcinoma cell metabolome and suppressed tumorigenesis［J］. Carcinogenesis, 2014, 35(10): 2264-2272. DOI: 10.1093/carcin/bgu174. ［8］ Luo H, Guo W, Wang F, et al. miR1291 targets mucin 1 inhibiting cell proliferation and invasion to promote cell apoptosis in esophageal squamous cell carcinoma［J］. Oncol Rep, 2015, 34(5): 2665-2673. DOI: 10.3892/or.2015.4206. ［9］ Tong D, Liu Q, Liu G, et al. The HIF/PHF8/AR axis promotes prostate cancer progression［J］. Oncogenesis, 2016, 5(12): e283. DOI: 10.1038/oncsis.2016.74. ［10］ Ma Q, Chen Z, Jia G, et al. The histone demethylase PHF8 promotes prostate cancer cell growth by activating the oncomiR miR125b［J］. Onco Targets Ther, 2015, 8: 1979-1988. DOI: 10.2147/OTT.S85443. ［11］ Wang Q, Ma S, Song N, et al. Stabilization of histone demethylase PHF8 by USP7 promotes breast carcinogenesis［J］. J Clin Invest, 2016, 126(6): 2205-2220. DOI: 10.1172/JCI85747. ［12］ Sun L, Huang Y, Wei Q, et al. Cyclin ECDK2 protein phosphorylates plant homeodomain finger protein 8 (PHF8) and regulates its function in the cell cycle［J］. J Biol Chem, 2015, 290(7): 4075-4085. DOI: 10.1074/jbc.M114.602532. ［13］ Li X, Gong X, Chen J, et al. miR340 inhibits glioblastoma cell proliferation by suppressing CDK6, cyclinD1 and cyclinD2［J］. Biochem Biophys Res Commun, 2015, 460(3): 670-677. DOI: 10.1016/j.bbrc.2015.03.088.

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