抑制Her-2基因的表达对NK-92细胞杀伤SKOV3细胞活性影响的研究

目的:探讨抑制Her-2基因的表达对NK-92细胞杀伤SKOV3细胞活性的影响。方法:Her-2siRNA质粒在脂质体介导下转染到包装病毒细胞株PT67中,将病毒上清转染SKOV3,经嘌呤霉素筛选得到稳定抑制Her-2基因表达的SKOV3/siRNA-1、SKOV3/siRNA-2细胞株,并分别通过RT-PCR和免疫组化法鉴定抑制Her-2表达的效果。应用LDH法检测NK-92细胞对SKOV3、SKOV3/siRNA-1、SKOV3/siRNA-2、SKOV3/siRNA-neg-ative control的杀伤活性。结果:Her-2/siRNA-1、Her-2/siRNA-2均可有效沉默Her-2mR-NA和蛋白的表达。NK-92细胞对SKOV3、SKOV3/siRNA-negative control的杀伤率分别为21%、20%,而对SKOV3/siRNA-1、SKOV3/siRNA-2的杀伤率分别为33%、45%(P<0·05)。结论:抑制Her-2基因的表达可增强NK-92细胞对SKOV3细胞的杀伤活性。RNA干扰技术联合NK-92细胞为高表达Her-2基因卵巢癌的生物治疗提供了一种新的策略。

Enhancement of cytotoxic activity of NK-92 cells against SKOV3 by inhibition of the expression of Her-2 gene

Objective:To explore the effects of inhibition of Her-2 gene expression on cytotoxic activity of NK-92 cells against SKOV3.Methods:The vectors-based siRNAi was transfected into packaging cell line PT67,SKOV3 was infected by the virus containing supernatant of stable PT67 cell lines,and the SKOV3/siRNA-1、SKOV3/siRNA-2 cell lines with a persistent knockdown of Her-2 gene were established by selection with puromycin,the reduction of Her-2 mRNA and p185 was investigated by RT-PCR and immumohistochemistry methods.The cytotoxic activity of NK-92 cells against SKOV3,SKOV3/siRNA-1,SKOV3/siRNA-2,SKOV3/siRNA-negative control was detected by LDH.Results:Both Her-2/siRNA-1 and Her-2/siRNA-2 could silence the expression of Her-2 gene.The cytotoxicity of NK-92 against SKOV3 and SKOV3/ siRNA-negative control was 21 and 20 respectively,while the cytotoxicity of NK-92 against SKOV3/siRNA-1 and SKOV3/siRNA-2 was 33 and 45(P<0.05).Conclusions:The cytotoxic activity of NK-92 cells against SKOV3 can be increased by inhibition the expression of Her-2 gene.NK-92 cytoxicity against tumor cells in combination with RNAi provide a new strategy for biological treatment of ovarian carcinoma with highly expression of Her-2 gene.