导入RASSF1A基因抑制膀胱癌细胞增殖的研究

Study on RASSF1A Gene Transfer for Inhibiting Cell Proliferation of Bladder Cancer Cell Line

[目的]探讨RASSF1A基因的表达对人膀胱癌细胞株体外增殖的影响.[方法]通过脂质体介导的基因转染法将野生型RASSF1A基因的真核表达载体及空载体转染膀胱癌BIU87细胞株.以四甲基偶氮唑蓝（MTT）法检测细胞的增殖程度,原位凋亡细胞检测技术和流式细胞仪检测膀胱癌BIU87细胞凋亡和细胞周期,Western blot检测膀胱癌BIU87细胞Cyclin D1蛋白表达的改变.[结果]成功建立稳定表达野生型RASSF1A基因的膀胱癌细胞株.野生型RASSF1A基因的表达可明显抑制膀胱癌细胞在体外的生长,可以显著降低BIU87细胞的增殖比和增高BIU87细胞的凋亡指数,与对照相组比差异显著（ P 〈0.05）.BIU87细胞的细胞周期发生 G1/S期阻滞,Cyclin D1蛋白的表达水平下调.[结论]野生型RASSF1A的重表达通过下调Cyclin D1,抑制膀胱癌BIU87细胞体外增殖.

[Objective]To explore the effect of RASSF1A gene expression on the proliferation of bladder cancer cell line in vitro. [Methods]The plasmids including pcDNA 3. 1 （＋）RASSF1A and pcDNA 3. 1 （＋） were separately transferred into bladder cancer＇cell line BIU87. MTT method was used to detect cell prolifera- tion. Tunnel and FCM were used to detect cell apoptosis and cycle of BIU87 cells. Western blot was used to detect the CyclinD1 protein expression in BIU87 cells. [Results] BIU87 cell lines that stably expressed wide- type or mutant RASSF1A gene were established successfully. The expression of wide-type RASSF1A gene could obviously inhibit the growth of bladder cancer cells in vitro, markedly reduce the proliferation ratio and increase the apoptotic index of BIU87 cells. There were significant differences between two groups （ P 0.05）. The cell cycle of BIU87 arrested at G1/S. The expression of CyelinD1 protein decreased. [Conclusion] Reexpression of wide-type RASSF1A can inhibit the proliferation of BUI87 cells in vitro by the down-regula- tion of CyclinD1 protein.