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  

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

作　　者：周颖[1] 凌斌[1] 姚凤球[1] 沈国栋[1] 冯定庆[1] 陈峥峥[1] 石永云[1] 高婷[1] 王梅梅[1] 祝怀平[1] 

机构地区：[1]安徽省分子医学重点实验室

出　　处：《现代妇产科进展》2006年第9期675-677,I0003,共4页Progress in Obstetrics and Gynecology

基　　金：安徽省自然基金课题(20050430715);安徽省教育厅(2005KJ347ZC)

摘　　要：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。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.

关 键 词：基因 Her-2 杀伤细胞 卵巢癌细胞株 RNA干扰 

分 类 号：R392.1[医药卫生—免疫学]