顺铂对人肺腺癌细胞作用与ERCC1、Bcl-2表达的相关性研究

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: 10 μg/mL DDP作用 12 h,A549细胞中ERCC1、Bcl-2 mRNA表达开始增高,随着作用时间的延长,ERCC1、Bcl-2 mRNA的表达逐渐增高,在 72 h 表达最强。浓度为 5 μg/mL 的DDP即可刺激A549细胞中ERCC1、Bcl-2 mRNA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 20 μg/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-2 mRNA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。

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

AIM: To study the relationship between the expressions of ERCC1, Bcl-2 and the effect of cisplatin in human lung adenocarcinoma cell lines,which suggested that the effects of ERCC1, Bcl-2 in cisplatin resistance effect.METHODS: A549 and A549/DDP cell lines were collected in the study,then the resistance of A549/DDP cells was measured by MTT assay.The expressions of ERCC1, Bcl-2 mRNA and protein after different concentrations' and different time' intervention were measured by immunocytohistology SABC assay and RT-PCR respectively.RESULTS: After treating with 10 μg/mL cisplatin for 12 h, the expressions of ERCC1, Bcl-2 mRNA and protein were increased in A549 cells; With the extension of time, the expressions of ERCC1 and Bcl-2 mRNA were increased gradually, they reached the peak levels in 72 h group.After treating with 5 μg/mL cisplatin for 24 h, the expressions of ERCC1, Bcl-2 mRNA and protein was increased in A549 cells.With the increase of concentration, the expressions of ERCC1 and Bcl-2 mRNA were increased gradually ,they reached the highest levels in 20 μg/mL group.Compared with the paremtal cells, the expressions of ERCC1 and Bcl-2 mRNA and protein were increased significantly in A549/DDP cells.CONCLUSION: With increasing concentrations of cisplatin and action time, the expressions of ERCC1, Bcl-2 mRNA and protein are increased in A549 cells,then speculates that ERCC1 and Bcl-2 may be involved in cisplatin acquired resistance.