Rab25基因siRNA诱导人卵巢癌细胞A2780……

 目的 观察Rab25基因的小分子干扰RNA（siRNA）对肿瘤细胞的诱导凋亡作用。方法 成功构建Rab25基因的siRNA，稳定转染靶细胞，MTT法检测细胞增生率；流式细胞仪（FCM）检测细胞生长周期和细胞凋亡率；DNA琼脂糖凝胶电泳法（DNA ladder）确定凋亡。结果 细胞生长明显受到抑制，增生能力下降；阻断细胞生长于G1期（转染pSUPER／Rab25 siRNA组细胞G1期达到88.6 %）；细胞凋亡增加，转染pSUPER／Rab25 siRNA组细胞凋亡率为12.3 %，与A2780组细胞及转染空载体组细胞相比，差异有统计学意义（P＜0.01）；DNA凝胶电泳可观察到细胞凋亡特异的DNA梯形带。结论 Rab25基因的siRNA可抑制对肿瘤细胞的生长，诱导其凋亡，可为卵巢癌基因治疗开辟新途径

Study on apoptosis of human ovarian cancer cell induced by siRNA on Rab25

Objective To study the effect of siRNA of Rab25 on apoptosis induction in ovarian carci－noma cell. Methods According to Rab25 mRNA sequence in the Genebank, the plasmids expressing siRNA against Rab25 were constructed and stably transected into A2780 cells, MTT method were applied to measure cell growth and proliferation. Apoptosis rate and cell cycle phase distribution of A2780 cells were measured by flow cytometry (FCM). Apoptosis was confirmed by agarose gel electrophoresis(AGE) of DNA. Results Cells transected with the plasmids expressing siRNA targeting Rab25 gene effectively decreased cell growth, proliferation; blocked A2780 cells in the G1 phase of cell cycle and induced cell apoptosis. A typical DNA ladder pattern appeared on AGE. Conclusion Rab25 gene siRNA can inhibit growth and induce apoptosis in ovarian carcinoma cell line,which will facilitate further studies of Rab25 function and its application in the treatment of ovarian cancer