MicroRNA-30a在膀胱癌中的表达及作用

doi:10.11904/j.issn.1002-3070.2017.04.005

摘要/Abstract

摘要： 目的探究微小RNA(miRNA)在人膀胱癌细胞系中的表达及其对人膀胱癌细胞增殖、凋亡及迁移侵袭能力的影响。方法应用荧光实时定量PCR(qRT-PCR)法检测膀胱癌细胞系(5637和T24)和膀胱上皮永生化细胞(SV-HUC-1)中miRNA-30a的表达水平。通过对T24细胞转染miR-30a mimic和5637细胞转染miR-30a inhibitor上调或下调miR-30a的表达,并对其分别转染NC mimic和NC inhibitor作为对照。利用流式细胞技术、MTT法和Transwell法探究miR-30a的表达对膀胱癌细胞增殖、凋亡以及侵袭能力的影响。结果在两种膀胱癌细胞系(5637和T24)中miRNA-30a的表达显著低于正常膀胱细胞系SV-HUC-1,并且在恶性度较高的T24膀胱癌细胞中其表达水平明显低于恶性度相对较低的5637细胞系。细胞转染72h后,miR-30a mimic组T24细胞OD值(0.83±0.09)明显低于NC mimic组(1.21±0.12)(P=0.003);miR-30a inhibitor组5637细胞OD值(1.28±0.14)高于NC inhibitor组(1.09±0.14)(P=0.019)。miR-30a mimic组T24细胞凋亡率(21.27±2.42)%明显高于NC mimic组(10.61±1.29)%;miR-30a inhibitor组5637细胞凋亡率(6.78±2.57)%明显低于NC mimic组(13.42±1.40)%,差异均具有统计学意义(P=0.0002,P=0.0014)。miR-30a mimic组穿膜细胞数(183.57±16.61)低于NC mimic组(465.80±9.20)(P＜0.0001);miR-30a inhibitor组(581.25±11.02)高于NC mimic组(397.13±7.57)(P＜0.0001)。结论 miR-30a表达的上调能够抑制膀胱癌细胞的增殖,促进细胞的凋亡,并降低膀胱癌细胞的迁移及侵袭的能力。膀胱癌细胞中miR-30a的低表达可能与膀胱癌的发生发展及转移有关。

关键词: 膀胱癌细胞, miRNA-30a, 增殖, 凋亡, 侵袭

Abstract: Objective The aims of this study were to investigate the expression of microRNA-30a(miR-30a)in human bladder cancer cell lines and their effects on the proliferation,apoptosis and migration of human bladder cancer cells.Methods The expression levels of miR-30a in bladder cancer cell lines(5637 and T24)and bladder epithelial immortalized cells(SV-HUC-1)were detected by real-time quantitative PCR(qRT-PCR).The expression of miR-30a was up-regulated or down-regulated by T24 cells transfected with miR-30a mimic or 5637 cells transfected with miR-30a inhibitors and controls using NC mimic or NC inhibitor.The effects of miR-30a expression on the proliferation,apoptosis and invasion of bladder cancer cells were investigated by flow cytometry,MTT and Transwell assays.Results The expression level of miR-30a in two bladder cancer T24 and 5637 cell lines was significantly lower than that in normal bladder SV-HUC-1 cell line(P＜0.05),and the expression level of miR-30a was lower in the high degree of malignancy in bladder cancer T24 cells than that in malignant degree of relatively low 5637 cells.After 72h transfection,the values of optical density(OD)in the miR-30a mimic group(0.83±0.09)was significantly lower than that in NC mimic group(1.21±0.12)in T24 cells(P＜0.01).The OD values of miR-30a inhibitor group(1.28±0.14)was significantly lower than that in the NC inhibitor group(1.09±0.14)in 5637 cells(P＜0.01).The apoptotic rate of miR-30a mimic group in T24 cells(21.27±2.42)% was significantly higher than that in the NC mimic group(10.61±1.29)%(P＜0.01).The apoptotic rate of the miR-30a inhibitor group in 5637 cells(6.78±2.57)% was significantly lower than that in the NC mimic group(13.42±1.40)%(P＜0.01).The number of transmembrane cells in miR-30a mimic group in T24 cells(183.57±16.61)was significantly lower than that in NC mimic group(465.80±9.20)(P＜0.01).The number of transmembrane cells in the miR-30a inhibitor group in 5637 cells(581.25±11.02)was significantly lower than that in NC mimic group(397.13±7.57)(P＜0.01).Conclusion Up-regulation of miR-30a can inhibit the proliferation of bladder cancer cells,promote cell apoptosis and reduce the ability of migration and invasion in bladder cancer cells.The low expression of miR-30a in bladder cancer cells may be related to the development and metastasis in bladder cancer.

Key words: Bladder cancer cell, miRNA-30a, Proliferation, Apoptosis, Invasion

中图分类号:

R737.14

柳清,尤泊森,马景光,邢丽娜. MicroRNA-30a在膀胱癌中的表达及作用[J]. 实用肿瘤学杂志, 2017, 31(4): 310-315.

LIU Qing,YOU Bosen,MA Jingguang,XING Lina. The expression and function of microRNA-30a in bladder cancer[J]. PRACTICAL ONCOLOGY JOURNAL, 2017, 31(4): 310-315.

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