RNA干扰survivin基因对人食管癌EC109细胞体外增殖和化疗敏感性的影响

Study on RNAi-mediated inhibition of survivin gene on proliferation and chemosensitivity of human esophageal cancer EC109 cells in vitro

目的研究RNA干扰survivin基因对EC109细胞体外增殖和化疗药物敏感性的影响。方法构建siRNA-survivin真核表达载体,转染EC109细胞,筛选得到稳定细胞克隆。同时设空载体和构建非特异siRNA载体转染为对照;MTT法检测各组细胞的体外增殖速度和比较它们对化疗药物顺铂的敏感性;流式细胞仪检测各组细胞的周期变化。结果siRNA-survivin转染的EC109细胞体外增殖受到抑制;不同浓度顺铂对各组细胞的增殖有抑制作用,且呈剂量依赖性。siRNA-survivin转染的EC109细胞的IC50为1.0188μg/mL,低于其他组(P<0.05);siRNA-survivin转染的EC109细胞组细胞周期明显阻滞于G0/G1期(P<0.05),S期比例也低于其他组(P<0.05)。结论RNA干扰survivin基因具有抑制人食管癌EC109细胞体外增殖和提高细胞对化疗药物顺铂敏感性的作用,将更多细胞阻滞在G0/G1期是其重要的作用机制。

[Objective] To investigate the RNA interference-mediated inhibition of survivin gene on the proliferation and chemosensitivity to cisplatin of EC109 cells in vitro. [Methods] siRNA-survivin eukaryotic expression vector was constructed to transfect EC109 cells which were furtherly screened and evaluated to select stable cell clones. Meanwhile, empty vector and non-specific siRNA vector transfection were prepared as controls. Proliferation in vitro of all groups of cells were measured and their sensitivity to cisplatin, a chemotherapy drug, compared by means of MTT assay. Cell cycles of all groups of cells were detected by flow cytometer. [Results] Proliferation in vitro of siRNA-survivin transfected EC 109 cells was suppressed. Cisplatin with different density can inhibit the proliferation of all groups of cells in a dose-dependent manner. IC50 of siRNA-survivin transfected EC109 cells was 1.0188 μmL, lower than that of control groups with a marked difference （P 〈0.05）. Cell cycle of the experimental group was obviously blocked in G0/G1 phase （P 〈0.05）, and the proportion of S period of this group was also lower than that of control groups （P 〈0.05）. [Conclusions] RNAi-mediated survivin gene can inhibit proliferation in vitro of human esophageal cancer EC109 cells, and improve the sensitivity of cells to cisplatin, a chemotherapy drug. Induction of G0/G1 cell cycle arrest might be its important mechanism.