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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 investigate the effect of RASSF1A gene expression on the apoptosis of human bladder cancer cells and its chemosensitivity. Methods The eukaryotic expression vector of wild-type and mutant RASSF1A gene and empty vector were transfected into BIU87 cell line by liposome-mediated gene transfection. The cell lines were divided into blank control group (group A) (Group B), transfected with mutant RASSF1A (group C) and wild type RASSF1A (group D). Groups B, C and D were treated with mitomycin C The proliferation of BIU87 cells was detected by azolyl blue staining and the apoptosis of bladder cancer BIU87 cells was detected by in situ apoptosis detection. Western blot assay Caspase-3 protein expression levels. RESULTS: The bladder cancer cell lines stably expressing wild-type and mutant RASSF1A genes were successfully established. Before and after adding mitomycin, the cell proliferation ratio, apoptosis index and the expression level of Caspase-3 in group D were significantly lower than those in group A and B (P <0.05). There was no significant difference between C and A, B groups (P> 0.05). Conclusion The overexpression of wild-type RASSF1A can promote the apoptosis of bladder cancer BIU87 cells and increase the sensitivity of bladder cancer cells to mitomycin.