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目的 观察生存素基因突变体质粒 (Cys84Ala)对胃癌细胞有丝分裂过程中细胞动力学行为的影响。方法 应用基因重组技术构建Cys 84Ala ,以脂质体转染的方法将质粒DNA转入胃癌细胞株 ,应用免疫组化法检测胃癌组织中生存素基因表达 ,流式细胞仪检测胃癌细胞凋亡及细胞周期 ,West ernblot检测胃癌细胞中多聚ADP核糖聚合酶 (PARP)和细胞色素C蛋白的表达 ,免疫荧光染色检测有丝分裂危象情况。结果 Cys84Ala可通过抑制野生型生存素基因的功能诱导胃癌细胞凋亡 ,增加cas pase 3活性 ,促进PARP裂解和细胞色素C的释放 ,亦可导致胃癌细胞发生有丝分裂障碍而死亡。 结论 Cys84Ala能通过不同的途径诱导胃癌细胞凋亡和有丝分裂危象 ,并有可能成为胃癌治疗的一个候选基因。
Objective To investigate the effect of survivin gene mutant plasmid (Cys84Ala) on the cellular dynamics of gastric cancer cells during mitosis. Methods Cys 84Ala was constructed by gene recombination technique. The plasmid DNA was transfected into gastric cancer cell lines by liposome transfection. The expression of survivin gene was detected by immunohistochemistry. The apoptosis of gastric cancer cells was detected by flow cytometry. Cell cycle, Western ernblot detection of gastric cancer cells ADP ribose polymerase (PARP) and cytochrome C protein expression, immunofluorescence staining mitotic crisis detection. Results Cys84Ala could induce the apoptosis of gastric cancer cells by inhibiting the function of wild-type survivin gene, increase the activity of caspase 3, promote the cleavage of PARP and the release of cytochrome C, and lead to the death of gastric cancer cells due to mitosis. Conclusion Cys84Ala can induce gastric cancer cell apoptosis and mitotic crisis through different pathways and may be a candidate gene for gastric cancer therapy.