荜茇酰胺对人肺癌A549/DDP细胞耐药性的逆转作用

目的:研究荜茇酰胺对人肺癌A549/顺铂(DDP)细胞耐药性的逆转作用。方法:A549/DDP细胞经0、20、30μmol/L荜茇酰胺作用48 h后,用MTS法检测肿瘤细胞抑制率;流式细胞术检测肿瘤细胞凋亡、细胞周期、P-糖蛋白(P-gp)表达和肿瘤细胞内罗丹明Rht123含量的变化;Western blotting法检测多药耐药基因(MDR)1、多药耐药相关蛋白(MRP)1、DNA拓扑异构酶(Top)Ⅱ、谷胱甘肽S-转移酶(GST)-π、凋亡抑制蛋白Survivin、周期蛋白依赖性蛋白激酶(CDK)1和蛋白激酶(PK)Cζ蛋白表达;实时荧光聚合酶链反应(RT-PCR)法检测MDR1、MRP1、Top-II、GST-π、Survivin和CDK1 m RNA表达;酶标仪检测含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-3、8活性。结果:A549/DDP细胞经0、20、30μmol/L荜茇酰胺作用48 h后,DDP对肿瘤细胞增殖的抑制率明显升高;与0μmol/L比较,20、30μmol/L荜茇酰胺作用48 h后,DDP导致的细胞凋亡率和G2期/M期明显升高,P-gp表达明显减弱,Rh-123浓度明显增加,MDR1、MRP1、Top-II、GST-π、Survivin、CDK1和PKCζ蛋白表达明显减弱,MDR1、MRP1、Top-Ⅱ、GST-π、Survivin、CDK m RNA表达明显减弱,Caspase-3、8的活性明显增强。结论:荜茇酰胺可逆转人肺癌A549/DDP细胞DDP耐药性,可能与其调节多药耐药相关基因表达有关。

Reversal Effects of Piperlongumine on Drug Resistance of Human Lung Caner A549/DDP Cell to Cisplatin

OBJECTIVE： To study the reversal effects of piperlongumine （PL） on drug resistance of human lung cancer A549/ DDP cell to cisplatin. METHODS： After treated with different concentrations of PL for 48 h, the inhibition rate of A549/DDP cell was determined by MTS assay. Cell apoptosis, cell cycle, P-gp expression and the content of Rh-123 were determined by flow cy- tometry. The protein expressions of MDR1, MRP1, Top-Ⅱ, GST-π, Survivin, CDK1 and PKCζ were detected by Western blotting assay. The mRNA expressions of MDR1, MRP1, Top-Ⅱ, GST-π, Survivin and CDK1 were detected by RT-PCR. The activities of Caspase-3 and Caspase-8 containing aminothiopropionic acid were detected by ELIASA. RESULTS： After treated with PL （0, 20, 30 μmol/L） for 48 h, inhibition rate of cisplatin to A549/DDP cell was increased significantly. Compared with 0 μmol/L PL, 20, 30 μmol/L PL could improve the rate of cisplatin-induced cell apoptosis and G2/M, the content of Rh-123 and the activities of Cas- pase-3 and Caspase-8 significantly, while decreased the expression of P-gp, the protein expression of MDR1, MRP1, Top-Ⅱ, GST-π, Survivin, CDK1 and PKCζ, mRNA expression of MDR1, MRP1, Top-Ⅱ , GST-π, Survivin and CDK1. CONCLU- SIONS： PL can reverse drug resistance of human lung cancer A549/DDP cell to cisplatin, which may be associated with the regula- tion of drug resistance-related gene expression.