MicroRNA-30a在膀胱癌中的表达及作用

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72h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.%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 72h 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.