论文部分内容阅读
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。
AIM: To investigate the effect of inhibiting the expression of Her-2 gene on the cytotoxicity of NK-92 cells to SKOV3 cells. Methods: Her-2 siRNA plasmid was transfected into packaging virus cell line PT67 by lipofectamine. The virus supernatant was transfected into SKOV3, and SKOV3 / siRNA-1 stably inhibited the expression of Her-2 gene , SKOV3 / siRNA-2 cell line, and the effect of inhibiting Her-2 expression was identified by RT-PCR and immunohistochemistry respectively. The killing activity of NK-92 cells on SKOV3, SKOV3 / siRNA-1, SKOV3 / siRNA-2, SKOV3 / siRNA-neg-ative control was tested by LDH method. Results: Her-2 / siRNA-1 and Her-2 / siRNA-2 all effectively silence Her-2mR-NA and protein expression. The killing rates of SKOV3 and SKOV3 / siRNA-negative control in NK-92 cells were 21% and 20%, respectively. The killing rates of SKOV3 / siRNA-1 and SKOV3 / siRNA-2 were 33% and 45% 0 · 05). Conclusion: Inhibition of Her-2 gene expression can enhance the cytotoxic activity of NK-92 cells to SKOV3 cells. RNA interference technology and NK-92 cells provide a new strategy for biotherapy of overexpressing Her-2 gene in ovarian cancer.