RNA干扰survivin基因在EC109细胞中作用的体内实验研究

A Study on RNAi-mediated Inhibition of survivin Gene on Proliferation of Human Esophageal Cancer EC109 Cells in Vivo

目的研究RNA干扰survivin基因对EC109细胞体内增殖的影响。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。同时设非特异siRNA载体转染为对照;通过裸鼠移植瘤模型的成瘤时间、瘤结节重量,比较各组细胞在体内的增殖速度;免疫组织化学染色检测移植瘤的细胞凋亡。结果siRNA-survivin转染的EC109细胞组在裸鼠体内成瘤时间为(8.86±1.069)d,高于其它各组;平均瘤重为(0.32±0.12)g,低于其它组,均有显著差异(P<0.05);TUNEL法细胞凋亡检测,siRNA-survivin转染的EC109细胞组的凋亡指数为(10.52±4.35),高于其它组,具有显著性差异(P<0.05);结论RNA干扰survivin基因具有抑制人食管癌EC109细胞体内增殖的作用,诱导细胞的凋亡是其重要作用机制。

Objective To investigate the RNA interference-mediated inhibition of survivin gene on the proliferation of EC109 cells in vivo. Methods siRNA-survivin eukaryotic expression vector was constructed to transfect EC109 cells which were further screened and evaluated to select stable cell clones. Meanwhile, non-specific siRNA vector transfection was prepared as control; Proliferation in vitro of three groups of cells were compared in terms of time of tumor formation and weight of tumor nodules on the model of implanted tumors in nude mice bearing esophageal cancer xenografts; Cell apoptosis of implanted tumors were detected by immunohistochemistry. Results Time of tumor formation of the group of siRNA-survivin transfected EC109 cells in nude mice was 8. 8571 ± 1. 06904 days, longer than that of control groups; mean tumor weight of the experimental group was 0. 3243 ± 0. 11998 g, lower than that of control groups, with a marked difference （P〈 0.05）; Cell apoptosis was detected by TUN-EL （TdT-mediated-biotinylated-dUTP nick end labeling）, with the finding that apoptosis index（AI） of the experimental group was 10.52 ± 4.35, higher than that of control groups, with a marked difference （P 〈 0.05） ; Conclusions RNAi-mediated survivin gene can inhibit proliferation in vivo of human esophageal cancer EC109 cells and the major mechanism is induce cell apoptosis.