CTR1和ATP7B在人肺腺癌细胞A549的表达及其与顺铂耐药的关系

Resistance to cisplatin is mediated by synergistic action of CTR1 and ATP7B in A549 cell line in vitro

摘要:

目的  探讨人肺腺癌细胞株A549和肺腺癌顺铂耐药细胞株A549/DDP对顺铂的耐药机制。方法  利用噻唑蓝法检测顺铂对A549/DDP及其亲代细胞株的细胞毒作用和生长曲线的影响，探讨A549/DDP细胞株凋亡敏感性的变化规律。采用Western blot检测该细胞株铜离子转运蛋白1（CTR1）、铜离子转运磷酸化ATP酶（ATP7B）及聚腺苷二磷酸-核糖聚合酶（PARP）剪切蛋白表达，分析A549细胞对顺铂耐药与CTR1、ATP7B及PARP剪切蛋白表达量的相关性。结果  A549/DDP较其亲代细胞株对顺铂引起的细胞毒性和凋亡敏感性下降；较其亲代细胞株A549/DDP中ATP7B表达增加，CTR1表达降低。结论  人肺腺癌细胞株A549对顺铂耐药的产生与细胞凋亡敏感性降低有关，且细胞内ATP7B表达增加，CTR1表达降低，或可作为细胞对顺铂的增敏靶点。

Abstract:

Objective To study the mechanisms of cisplatin resistance of human lung adenocarcinoma cell lines A549 and A549/DDP. Methods Human lung adenocarcinoma A549 cells and a cisplatin-resistant derivative A549/DDP were treated with varying concentrations of cisplatin. The changes in cell growth, cytotoxicity and apoptosis were explored. Western blot was used to determine the expression of copper transporter 1 (CTR1), copper transporting phosphorylated ATPase (ATP7B) and poly-ADP-ribose polymerase (PARP). The correlations of cisplatin resistance with CTR1, ATP7B and PARP were analyzed. Results Compared to the A549 cells, the A549/DDP cells were less sensitive to cisplatin-induced cytotoxicity and apoptosis. ATP7B expression increased but CTR1 expression decreased in the A549/DDP cells compared to the A 549 cells. Conclusions Our results demonstrate that reduced expression of the copper influx transporter CTR1 and overexpression of the copper efflux transporter ATP7B are responsible for the resistance of A549/DDP cells to cisplatin. These results indicate well that the expression of ATP7B and CTR1 may provide markers for chemoresistance and chemosensitivity, respectively, to cisplatin-based chemotherapy.