【摘要】 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。更多还原

【Abstract】 OBJECTIVE: To silence the expression of multidrug resistance(MDR1) gene in SGC7901/DDP cell line by using lentivirus mediated RNAi,so as to reverse its resistance to DDP.METHODS: The most effective interfering fragment targetting MDR-1 which was obtained by previous experiment,was packed by lentivirus expression vector and transfected into SGC7901/DDP cells.The interfering effect was proved by the methods of RT-qPCR and western blotting assay,and tests of cell’s drug sensitivity to DDP and apoptosis detection were performed.RESULTS: Lentivirus mediated gene silencing was far more effective than previous interfering plasmid.MDR-1 mRNA expression descended by(51±3)%,moreover.P-glycoprotein expression was down regulated to(47±8)%.IC50 of SGC-7901/DDP declined to 1.1 μg/mL after RNAi was carried out,and apoptosis rate was obviously upgraded.CONCULSION: Lentivirus mediated RNAi can effectively inhibit expression of MDR-1 in SGC-7901/DDP cell line,and reverse its drug resistence to DDP.更多还原