摘要
目的:建立Nrf2低表达的H460细胞株,可增敏奥沙利铂对肿瘤细胞的抗增殖作用,同时为Nrf2-ARE信号通路的研究提供便捷途径。方法:H460细胞转染pRS-hNrf2后经过筛选得到多个单克隆细胞株,通过Western blotting和GSH含量的检测方法分析各细胞株中Nrf2及其调控基因的表达以及GSH含量的变化,用MTS法检测细胞株对抗癌药物的敏感性。结果:H460细胞转染pRS-hNrf2后筛选建立Nrf2低表达的稳定细胞株H460-C9和H460-C13,与对照组相比其GSH含量均有显著降低(PC9=0.00249,PC13=0.03944);同时,MTS检测表明,抗癌药物奥沙利铂和阿霉素的抗细胞增殖作用在H460-C9和H460-C13细胞株中明显增强。结论:成功建立Nrf2低表达的H460-C9和H460-C13细胞株,与对照组细胞株相比,低表达Nrf2可增敏奥沙利铂对肿瘤细胞的抗增殖作用。
Objective: To establish a stable H460 cell line with Nrf2 down-regulation to study the role of Nrf2 in Oxaliplatin resistance.Methods: Nrf2 down-regulated H460 cell line was obtained by transfecting cells with pRS-hNrf2 and followed by screening for Nrf2 expression by Western blotting.The cell lines were further characterized by analysing cellular GSH levels and cytotoxicity to anti-cancer drugs with MTS.Results: Nrf2 down-regulated H460 cell lines were established successfully.Cellular GSH level reduced significantly in the H460-C9 and H460-C13 compared to control cells(PC9= 0.00249,PC13= 0.03944).Down-regulation of Nrf2 in H460 sensitized the anti-cancer effect of Oxaliplatin and Doxorubicin.Conclusion: Nrf2 plays an important role in drug resistance.Down regulation of Nrf2 in H460 cell enhances cytotoxicity of Oxaliplatin.
出处
《浙江大学学报(医学版)》
CAS
CSCD
北大核心
2010年第1期11-16,共6页
Journal of Zhejiang University(Medical Sciences)
基金
国家自然科学基金(30973555)
卫生部/浙江省卫生厅(WKJ2007-2-008)
浙江省科技厅(2008C23054)