MDR1及MDR3基因与卵巢癌细胞顺铂耐药的关系

目的:探讨MDR1(Multidrug resistance gene-1)及MDR3(Multidrug resistance gene-3)基因与卵巢癌细胞顺铂耐药的关系。方法:两种质粒一起瞬时转染A2780/DDP细胞,空载体组及未转染组作为对照。Annexin V-FITC/PI双标流式细胞术、MTT法、RT-PCR、western blot分别检测A2780/DDP细胞早晚期凋亡、细胞增殖活性、MDR1和MDR3 mRNA表达及caspase-3蛋白水平。结果:两种质粒(pSuppressorNeo MDR1 siRNA,MDR3 siRNA)均未能逆转A2780/DDP细胞顺铂耐药:①流式显示转染MDR1 siRNA组、MDR3 siRNA组和pSuppressorNeo空载体组后相比较未转染A2780/DDP细胞凋亡率无显著改变(P>0.05);②在一定药物浓度范围内,转染MDR1和MDR3小分子干扰RNA均未能增强顺铂对A2780/DDP细胞的细胞毒作用;③与空载体组和未转染细胞相比,A2780/DDP细胞的MDR3及MDR1 mRNA表达均低下(P>0.05);④Caspase-3蛋白表达未见增加。结论:顺铂所致的卵巢癌细胞多药耐药与MDR1及MDR3基因过度表达均无关,卵巢癌细胞顺铂耐药为非P-gp

The study of MDRI and MDR3 gene and drug resistant to cisplatin of ovarian cancer cells

Objective:To investigate the relationship between MDR1 and MDR3 gene and drug resistant to cisplatin of ovarian cancer cells.Methods:Two siRNAs(MDR1,MDR3) which were specifically targeted MDR1 and MDR3 gene were transferred into A2780/DDP cells.Then double stain with Annexin-V-FITC/PI(fluorescein isothiocyanate conjugated) to detect cell apoptosis by the flow cytometry(FCM).A2780/DDP cell viability was determined via MTT method.MDR1 and MDR3 mRNA were assessed by RT-PCR.Caspase-3 protein was detected by Western blot.Results:Transecting MDR1 and MDR3 siRNA into A2780/DDP cells could not result in different levels of reversal of drug-resistance to cisplatin(P>0.05).No a significant difference in the apoptosis efficiency was observed between the MDR1 and MDR3 siRNA,pSuppressorNeo vector transfection cells and untreated cells(P>0.05);In the presence of different concentrations of cisplatin,A2780/DDP cells viability was not significantly decreased after transfection.The expression of MDR1 and MDR3 mRNA was at low level in A2780/DDP cell,and the expression of MDR1 and MDR3 mRNA was not restrained by the MDR1 and MDR3 siRNA.The expression of caspase-3 protein of MDR1 and MDR3 siRNA transected group in A2780/DDP cells was not significantly increased.Conclusion:Multidrug resistance induced by cisplatin in ovarian carcinoma cell lines is not due to overexpression of MDR1 and MDR3 gene.Ovarian carcinoma cell cisplatin-dmg is a non-P-gp mediated MDR.