miR-195-5p稳定表达的膀胱癌T24细胞系的构建及其功能鉴定

Constructing and functional analysis of stable miR-195-5p-expressed T24 cell lines

目的:构建miR-195-5p稳定表达的膀胱癌T24细胞系,探讨miR-195-5p在膀胱癌T24细胞中的表达及高表达miR-195-5p对细胞生长的影响。方法:Real-time qRT-PCR(quantitative reverse tran-scription PCR)方法检测miR-195-5p在膀胱癌5637、T24、SW780细胞中的表达。通过构建pSilencer-miR-195-5p重组质粒,将其稳定转染至T24细胞,筛选miR-195-5p稳定表达细胞系。应用MTT实验和流式细胞术分别检测T24细胞高表达miR-195-5p对细胞增殖、凋亡的影响。结果:miR-195-5p在3种膀胱癌细胞系中普遍低表达,稳定转染的T24细胞具有miR-195-5p高表达特性,与对照细胞相比,其增殖活力和抗凋亡能力明显减低。结论:成功构建了能稳定表达miR-195-5p的T24细胞系,miR-195-5p高表达可以抑制细胞增殖、促进细胞凋亡。miR-195-5p低表达可能参与膀胱癌发生发展。

Objective：To construct stable miR - 195 -5p - expressed T24 cell lines and investigate the expression of miR - 195 -5 p in bladder cancer cells and its effect on cell growth. Methods：The expression of miR - 195 -5p in bladder cancer cells -5637 ,T24 and SW780 was determined using real -time quantitative reverse transcription PCR （ qRT - PCR ）. miR - 195 - 5 p expression vector was constructed and stably transfected into T24 ceils. The cell growth and apoptosis rate were evaluated in the stable T24 cells using M3T and flow cytometry assays. Results：miR - 195 -5p was down -regulated in 5637 ,T24 and SW780 cells compared with that in HUC cells. Stable expression of miR - 195 -5p in T24 cells induces a decrease of cell proliferation and an increase of apoptosis. Conclusion：The stable miR - 195 - 5p - expressed T24 cell lines were successfully established. Up - regulation of miR - 195 - 5p inhibits cell proliferation and promotes apoptosis, and the aberrant decreased expression of miR- 195 -5p may contribute tO bladder carcinogenesis.