CIK细胞对卵巢癌化疗耐药的逆转作用及对ERCC表达的影响

The reversal effect of CIK cells on chemoresistance in ovarian cancer and the effect on ERCC expression

目的探讨细胞因子诱导的杀伤(CIK)细胞在杀伤人卵巢癌耐顺铂(DDP)细胞株(SKOV3/DDP)的效应当中是否存在着逆转化疗耐药作用及其对切除修复交叉互补基因组1(ERCC1)、切除修复交叉互补基因组2(ERCC2)表达的影响。方法四甲基偶氮唑盐比色法(MTT)检测DDP、CIK细胞、无毒剂量CIK细胞联合顺铂对人卵巢癌亲本细胞株(SKOV3)和SKOV3/DDP的体外杀伤活性。实时荧光定量聚合酶链反应(real-time PCR)和Western-blot分别检测经无毒剂量CIK细胞作用下两组细胞中ERCC1、ERCC2基因表达的变化。结果 DDP和CIK细胞对SKOV3、SKOV3/DDP两组细胞均有杀伤效应,且随DDP浓度和效靶细胞比值的升高呈依赖性增加;与单纯DDP作用相比,无毒剂量CIK细胞联合DDP对SKOV3/DDP细胞具有较明显的协同作用(P<0.05)。CIK细胞作用SKOV3/DDP细胞后,靶细胞中ERCC1mRNA和蛋白的表达量较干预前降低,差异有统计学意义(P<0.05);ERCC2mRNA和蛋白的表达量较干预前降低,但差异无统计学意义(P>0.05)。结论 CIK细胞可以部分逆转SKOV3/DDP细胞对DDP的耐药性,且CIK细胞逆转SKOV3/DDP的耐药作用可能与ERCC1表达下调有关联,而与ERCC2表达下调无关联。

Objective To investigate the anti-tumor effect of cytokine induced killer(CIK)cells against cisplatin(DDP)resistant human ovarian cancer cell line(SKOV3/DDP),and its effect on the expression of excision repair cross-complementation genome 1(ERCC1)and excision repair cross-complementation genome 2(ERCC2).Methods The cytotoxic effects of DDP,CIK,non-toxic CIK combined with DDP against human ovarian cancer parental cell lines(SKOV3)and SKOV3/DDP were detected by MTT assay in vitro.Real-time PCR and Western-blot were used to detect the expression of ERCC1 and ERCC2 genes in the two groups of cells treated with non-toxic CIK.Results DDP and CIK had cytotoxic effects on both SKOV3 and SKOV3/DDP cells,in ratio of DDP concentration and target cells increased dependence.Compared with DDP alone,non-toxic CIK combined with DDP had a significant synergistic effect on SKOV3/DDP cells(P<0.05).The expression of ERCC1 mRNA and protein in SKOV3/DDP cells were lower than those before intervention(P<0.05).The expression of ERCC2 mRNA and protein were lower,but the difference was not statistically significant(P>0.05).Conclusion CIK cells can partially reverse the drug resistance of SKOV3/DDP cells to DDP,and it may be related to the down-regulation of ERCC1 expression but not to down-regulation of ERCC2 expression.