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目的:利用慢病毒介导的RNA干扰技术(RNAi),沉默SGC-7901/DDP胃癌耐药细胞株的多药耐药基因1(MDR-1)表达,逆转该细胞株对顺铂(DDP)的耐药性。方法:将前期筛选获得的MDR-1基因最佳干扰片段构建慢病毒表达载体,包装成慢病毒,感染SGC-7901/DDP耐药株,通过逆转录实时荧光定量PCR(RT-qPCR)和蛋白质印迹法验证干扰效果后进行DDP药物敏感度实验和细胞凋亡检测。结果:慢病毒介导的基因沉默效果较前期干扰质粒效果更明显,MDR-1mRNA表达抑制率为(51±3)%,且P糖蛋白表达下调至(47±8)%,干扰后的SGC-7901/DDP细胞IC50值下降至1.1μg/mL,并且细胞株的凋亡率显著增加。结论:慢病毒介导的RNAi可有效抑制SGC-7901/DDP胃癌耐药细胞株的MDR-1基因表达,实现其DDP耐药性逆转。
Objective: To silence the multidrug resistance gene 1 (MDR-1) expression in SGC-7901 / DDP multidrug resistance cell line by lentivirus-mediated RNA interference (RNAi) Drug resistance. Methods: The lentiviral expression vector was constructed by screening the best MDR-1 gene fragment and then packaged into lentivirus and infected with SGC-7901 / DDP drug-resistant strains. RT-qPCR and protein The DDP drug susceptibility test and apoptosis assay were performed after the interference effect was verified by the blotting method. Results: The effect of lentivirus-mediated gene silencing was more obvious than that of the previous plasmids. The inhibition rate of MDR-1 mRNA expression was (51 ± 3)%, and the expression of P-glycoprotein was down-regulated to (47 ± 8)% The IC50 of -7901 / DDP cells decreased to 1.1μg / mL, and the apoptosis rate of cell lines increased significantly. CONCLUSION: Lentivirus-mediated RNAi can effectively inhibit the MDR-1 gene expression in SGC-7901 / DDP gastric cancer cell line and reverse its DDP resistance.