腺病毒介导Her2／neu siRNA对卵巢癌细胞的生长抑制作用

Inhibition of tumor growth of ovarian cancer by siRNA mediated adenovirus targeting to Her2/neu gene

目的观察腺病毒介导的针对Her2／neu基因RNAi表达载体对Her2／neu的抑制及抑制后对卵巢癌SKOV-3细胞的生长的影响。方法通过构建的针对Her2neu的siRNA重组腺病毒（滴度为1．6×10^8PFU／mL）感染卵巢癌细胞SKOV-3后，采用Western—blot法观察Her2／neu基因的沉默效果，流式细胞仪分析细胞周期变化，cell Proliferation法检查细胞体外增殖能力。结果构建的重组腺病毒Adeno—Her2siRNA感染卵巢癌细胞SKOV-3后Her2／neu的蛋白表达降低；S期细胞较对照组未感染细胞比例增加；肿瘤细胞胞增殖速度缓慢（P〈0．05）。结论成功构建的重组腺病毒Adeno-Her2siRNA感染后可有效降低Her2／neu蛋白水平表达，阻滞细胞感染于S期，造成感染细胞生长速度减慢，有望为卵巢癌的基因治疗提供一种选择手段。

Objective To observe the Her2siRNA mediated adenovirus down-regulate Her2/neu gene and study the RNAi effect on the cell cycle and tumor growth of ovarian cancer SKOV-3 cells. Methods The ovarian cancer SKOV-3 cells was infected by the recombinant adenovirus vector Adeno-Her2siRNA（the titer was 1.6×10^8 PFU/mL）, the protein was detected by Western blot. FCM analysis and cell proliferation method were applied to measure cell cycle and cell growth respectively. Results The results from infection of SKOV-3 cells with siRNA-expressing adenovirus demonstrated that adenovirus-mediated RNAi could downregulate Her2/neu expression efficiently through Western blot. It can increase the number of cells in S phase, and inhibit the tumor cell growth （P〈0.05）. Conclusions The adenovirus-mediated Her2siRNA could effectively silence Her2/neu gene, increase the number of cells in S phase, and then slow down the growth of tumor cells. This may be a useful therapeutic strategy for ovarian cancer over-expressing Her2/neu.