CIK逆转耐顺铂人肺腺癌细胞对顺铂耐药与ERCC1的相关性

Study on the correlation between ERCC1 and the merchanisms underlying cytokine induced killer cells-reversed cisplatin resistance in A549/DDP Cells

目的观察脐血来源的CIK细胞对A549/DDP细胞的耐药逆转作用。并探讨CIK逆转耐药机制是否与核苷酸切除修复交叉互补基因1(excision repair cross complementation group 1,ERCC1)的表达存在相关性。方法采集脐血单个核细胞经细胞因子体外诱导获得CIK细胞(cytokine induced killer cells,CIK),MTT法检测顺铂(cisplatin,DDP)对A549细胞和A549/DDP细胞的体外杀伤作用,以及CIK细胞与A549/DDP细胞共培养前后对DDP敏感性的变化。Real-time RT-PCR法检测A549细胞、A549/DDP细胞以及CIK干预后的A549/DDP细胞中ERCC1 mRNA的表达情况。Western blot法检测A549细胞、A549/DDP细胞以及CIK干预后的A549/DDP细胞中ERCC1蛋白的表达情况。结果 A549/DDP细胞对DDP耐药,耐药指数为2.391。CIK细胞能逆转A549/DDP细胞对DDP的耐药性,逆转倍数为3.967。Real-time RT-PCR、Western blot测得A549/DDP细胞中ERCC1 mRNA和蛋白表达量比A549细胞高(P<0.05),CIK细胞共培养前后A549/DDP细胞中ERCC1 mRNA和蛋白表达量无明显变化。结论 CIK细胞能逆转A549/DDP细胞对DDP的耐药性,其逆转耐药的机制与ERCC1的表达无明显相关性。

Objective To investigate the effects of cord blood-derived cytokine-induced killer（CIK） cells on reversing cisplatin resistance in the DDP-resistant human lung adenocarcinoma cell line A549/DDP and observe whether there was correlation between these effects and excision repair cross complementation group 1（ERCC1）expression.Methods The single umbilical cord blood mononuclear cell was induced by cytokine in vitro to become CIK cells.MTT assay was applied to evaluate the cytotoxic activity of DDP against A549/DDP cells and A549 cells in vitro and the sensitivity of A549/DDP cells CIK to DDP before and after the co-culture with CIK.The mRNA and protein expressions of ERCC1 in A549 cells,A549/DDP cells and A549/DDP co-cultured with CIK cells were detected by real-time RT-PCR and western blot analysis.Results The A549/DDP cells was resistant to DDP with the resistance index of 2.391.CIK cells decreased drug resistance to cisplatin with the reversal fold of 3.967.The ERCC1 mRNA and protein exprossions were higher in A549/DDP than those in A549cells（P 〈 0.05）,whereas there was no significant difference of the ERCC1 mRNA and protein exprossions in A549/DDP before and after A549/DDP co-cultured with CIK cells.Conclusion CIK could reverse the resistance of A549/DDP cells to DDP and there was no significant correlation between the mechanisms and ERCC1 expression.