中文摘要：

目的：探讨AKT2基因在肿瘤细胞对紫杉醇敏感性中的作用。方法：分别构建2个针对同一AKT2基因不同位点的短发卡状RNA（shRNA）载体，转染人卵巢癌细胞A2780、SKOV3，荧光显微镜观察细胞内荧光蛋白的表达及转染效率，运用RT—PCR、蛋白质免疫印迹（western blotting）方法比较单独转染的抑制瘤和共转染对AKT2表达的沉默效率；抑制AKT2表达后，流式细胞仪（FACS）检测肿瘤细胞对紫杉醇的敏感性。结果：构建的2种shRNA表达载体均可抑制AKT2mRNA和蛋白的表达，共转染的抑制率明显高于分别单独转染的抑制率（P〈0．05）；FACS结果显示，共转染沉默AKT2基因后，细胞凋亡率明显增加（P〈0．05）。结论：共转染针对AKT2基因不同位点的2个shRNA真核表达载体，对AKT2基因的抑制效率高于单独转染1个载体；抑制卵巢癌细胞中AKT2基因表达，能增强其对紫杉醇的敏感性。

英文摘要：

Objective： To investigate the effect of AKT2 gene expression on the sensitivity of ovarian carcinoma cells to paclitaxel. Methods ： Vectors containing shRNA targeting two mutations of the same AKT2 gene were designed and constructed, respectively. The recombinants containing different AKT2 target sequences were transfected into human ovarian carcinoma cell lines A2780 and SKOV-3 cells, respectively. The efficiency of transfection was evaluated by fluorescent microscope. The reverse transcription-polymerase chain reaction （RT-PCR） and western blot were used to compare the efficiency of gene silencing between single transfection and co-transfection; the sensitivity of ovarian carcinoma to paclitaxel was evaluated by FACS after inhibition of AKT2. Results： Transfection of the two vectors inhibited the expression of AKT2 mRNA and protein in both A2780 and SKOV-3 cells. The gene silencing efficiency of co-transfection was significantly higher than that of each single transfection. The results of FACS indicated that apoptotic rate of ovarian cacinoma cells was obviously increased after silencing AKT 2 by co-transfection. Conclusion：Co-transfection of two different shRNA vectors against AKT2 gene has higher silencing efficiency than each single transfection of any one of the two vectors. Inhibition of AKT2 gene expression could sensitize ovarian carcinoma cells to paclitaxel.