生存素基因突变体诱导胃癌细胞有丝分裂障碍的实验研究

Experimental study on mitotic catastrophe induced by pcDNA_3-survivin-mutant in gastric cancer cell lines

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 Through gene reconstruction we generated pcDNA 3-survivin-mutant(Cys84Ala)plasmid, then the plasmid DNA was further transfected into gastric carcinoma cells by liposomal delivery. The effects of mutant survivin on cytokinetics of the gastric carcinoma cells were obversed. Methods By using immunohistochemical staining, the expression of survivin was detected in the gastric cancer tissues, and apoptosis was detected by flow cytometry. The PARP and cytochrome C expressions were determined by Western blot, the mitotic catastrophe was determined by immunofluorescence. Results Inhibition of survivin by mutant survivin cDNA could induce apoptosis, increase caspase-3 activity, cleave PARP and promote cytochrome C release in gastric cancer cells. Inhibition of survivin also caused mitotic catastrophe in gastric cancer cells. Conclusion Inhibition of survivin may induce apoptosis and mitotic catastrophe in gastric cancer. Survivin targeting gastric cancer therapy might be of potential benefit in the future.