miR-206对膀胱癌细胞生长的影响及分子作用机制研究

Effect of miR-206 on the growth of bladder cancer cells and its mechanism

目的 探讨miR-206对膀胱癌细胞T24和5637生长的影响及分子作用机制.方法 根据对膀胱癌细胞的不同处理,分为对照组（转染miR-NC）和实验组（转染miR-206）.EdU增殖实验和集落形成实验检测转染后膀胱癌细胞的增殖能力.流式细胞术检测膀胱癌细胞转染后的细胞周期分布.荧光实时定量PCR（qRT-PCR）和Western Blot检测CDK4和GAK表达水平的变化.结果 EdU增殖实验显示转染miR-206的膀胱癌细胞的增殖能力明显下降.集落形成实验显示,相比转染miR-NC组,miR-206组膀胱癌细胞形成的集落数数量更少.流式细胞术结果显示,转染miR-206后膀胱癌细胞在S期和G2/M期的细胞比例明显下降,而在G0/G1期的细胞比例明显升高.qRT-PCR和Western Blot结果显示,相比miR-NC,转染miR-206后T24和5637中CDK4和GAK表达水平均明显下降（P＜0.01）.结论 miR-206可显著抑制膀胱癌细胞的生长,其机制为miR-206干扰CDK4和GAK的表达,抑制膀胱癌细胞的细胞周期进展.

Objective To investigate the effect of miR-206 on the growth of bladder cancer cell lines T24 and 5637 and its molecular mechanism.Methods According to the different treatment,bladder cancer cells were divided into control group （transfected with miR-NC） and experimental group （transfected with miR-206）.EdU proliferation assay and colony-forming assay were performed to detect the proliferation ability of bladder cancer cells after transfection.Flow cytometry was used to detect the cell cycle distribution of bladder cancer cells after transfection.The expression of CDK4 and GAK was detected by real-time quantitative PCR （qRT-PCR） and Western Blot.Results EdU proliferation assay showed that the proliferation ability of bladder cancer cells transfected with miR-206 was significantly decreased.The colony formation assay showed that the number of colonies formed by the miR-206 transfected bladder cancer cells was lower than that of the miR-NC.The results of flow cytometry showed that the percentage of cells in S and G2 / M phase decreased significantly after miR-206 transfection,but the percentage of cells in G0 / G1 phase was significantly increased.The results of qRT-PCR and Western Blot showed that expressions of CDK4 and GAK in T24 and 5637 significantly decreased after transfection of miR-206 compared to miR-NC （P ＜ 0.01）.Conclusions miR-206 significantly inhibits the growth of bladder cancer cells.The mechanism is that miR-206 inhibits the cell cycle progression of bladder cancer cells by interfering the expressions of CDK4 and GAK.