顺铂对人肺腺癌细胞作用与ERCC1、Bcl-2表达的相关性研究

Relationship between ERCC1,Bcl-2 expression and cisplatin intervention in human lung adenocarcinoma cell lines

目的:探讨ERCC1、Bcl-2表达与顺铂作用的关系,进而推测ERCC1、Bcl-2在顺铂耐药中的作用。方法:选择人肺腺癌细胞A549及其耐DDP细胞株A549/DDP作为研究对象,采用MTT法检测A549/DDP细胞耐药指数;免疫细胞化学方法、RT-PCR方法检测不同浓度、不同作用时间干预后细胞中ERCC1、Bcl-2的表达。结果:10μg/mL DDP作用12h,A549细胞中ER-CC1、Bcl-2mRNA表达开始增高,随着作用时间的延长,ERCC1、Bcl-2mRNA的表达逐渐增高,在72h表达最强。浓度为5μg/mL的DDP即可刺激A549细胞中ERCC1、Bcl-2mRNA表达水平上升,随着DDP浓度的增加,其表达水平逐渐上升,ERCC1民mRNA表达水平在20μg/mL组达到高峰。与亲本细胞相比,A549/DDP中ERCC1、Bcl-2表达明显增高。结论:随着顺铂作用时间的延长和浓度的增加,A549细胞中ERCC1、Bcl-2mRNA和蛋白的表达量增加,推测ERCC1、Bcl-2可能参与了顺铂继发耐药的形成。

Objcetive The excision repair cross-complementing gene 1（ERCC1） and Bcl-2 are reported to be related to cisplatin resistance in tumor cells.The aim is to confirm the relationship between ERCC1,Bcl-2 and cisplatin resistance in human lung adenocarcinoma cell lines. Methods A549 and A549/DDP cell lines were treated with 10ug/ml cisplatin for 12,24,48,72h,or treated with 5,10,20,40 ug/ml cisplatin for 24h respectively. Then the resistance of A549/DDP cells was measured by MTT assay. The expression of ERCC1,Bcl-2 mRNA and protein was measured by immunocytohistology SABC assay and PR-PCR respectively. Results After treating with 10ug/ml cisplatin for 12h, up-regulation of ERCC1,Bcl-2 mRNA and protein was observed in A549 cells; Then reached the peak levels in 72h group. After treating with 5ug/ml cisplatin for 24h, up-regulation of ERCC1, Bcl-2 mRNA and protein was observed in A549 cells.When treated with 20ug/ml cisplatin for 24h,the ERCC1 mRNA and protein reached the peak levels. Comparing with the parental cells, ERCC1and Bcl-2 expression increased obvious in A549/DDP cells. Conclusion Up-regulation of ERCC1, Bcl-2 exprission can be induced by low-dose cisplatin in human lung adenocarcinoma cell line A549,and ERCC1and Bcl-2 may play roles in cisplatin acquired resistance.