RASSF1A基因对膀胱癌细胞凋亡及化疗药物敏感性的影响

Effects of RASSF1A expression on chemotherapy sensitivity and apoptosis of bladder cancer cell

目的探讨RASSF1A基因表达对人膀胱癌细胞凋亡及对化疗药物敏感性的影响。方法采用脂质体介导的基因转染法建立野生型和突变型RASSF1A基因的真核表达载体及空载体转染膀胱癌BIU87细胞株,细胞株分为空白对照组(A组)、空载组(B组)、转染突变型RASSF1A组(C组)与转染野生型RASSF1A组(D组),B,C,D组采用化疗药物丝裂霉素作用于细胞,四甲基偶氮唑蓝法检测细胞增殖程度,原位凋亡细胞检测技术检测膀胱癌BIU87细胞凋亡。Western blot测定Caspase-3蛋白的表达水平。结果成功建立稳定表达野生型和突变型RASSF1A基因的膀胱癌细胞株;加入丝裂霉素前、后D组细胞增殖比、凋亡指数及Caspase-3蛋白表达水平与A,B组比较差异均有统计学意义(P<0.05),C组与A,B组比较,差异均无统计学意义(P>0.05)。结论野生型RASSF1A的重表达可促进膀胱癌BIU87细胞凋亡,提高膀胱癌细胞对丝裂霉素的敏感性。

Objective To study the effect of RASSF1A on chemotherapy sensitivity and apoptosis of bladder cancer cell. Methods The RASSF1A （wild-type or mutant） expression vector and empty vector were transferred into BIU87 cell lines. The BIU87 cell lines were divided into control group （group A）, transfeeted with empty vector group （group B）, transfected with mutant RASSFIA group （group C）, and transfected with wild-type RASSF1A group （group D）. The cells in groups B, C and D were treated with the mitomycin. MTT was used to detect the proliferation of BIU87 cell. Tunel was used to detect the apoptosis and Western blot was used to detect caspase-3 protein expression. Results The BIU87 cell lines stably expressing wide-type or mutant RASSF1A gene were established successfully. The expression levels of cell proliferation, apoptosis index and caspase-3 protein were significantly different in group A and B from those in group D （P〈0. 05）, and were not significantly differen from those in the group C （P〉0.05） both before and after being treated with the mitomycin. Conclusion Wild-type RASSFIA expression could induce BIU87 cells apoptosis and increase chemotherapy sensitivity of BIU87 cells to mitomycin.