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目的筛选靶向人胃癌SGC7901/VCR细胞多药耐药基因1(MDR1)小分子干扰RNA(siRNA)的有效序列。方法设计并体外转录靶向MDR1的4条siRNA(MDR1si326、MDR1si1513、MDR1si2631和MDR1si3071),转染SGC7901/VCR细胞。用RT PCR检测MDR1mRNA表达,免疫印迹检测P糖蛋白(P gp)表达,流式细胞仪检测细胞内阿霉素(ADR)蓄积,四甲基偶氮唑蓝(MTT)法检测细胞对ADR的敏感性。结果4条siRNA均能不同程度逆转SGC7901/VCR细胞MDR1介导的多药耐药。转染后48h,MDR1si326组和MDR1si2631组的mRNA表达水平分别为0.42±0.07和0.49±0.02,较MDR1si1513或MDR1si3071组明显下降(P<0.05);MDR1si326组细胞内ADR蓄积最显著,中位数为30.03,MDR1si2631组次之,MDR1si3071组较少,MDR1si1513组最少(P<0.05);MDR1si2631组对ADR耐药的相对逆转率最高,为(98.12±0.26)%,MDR1si326组次之(P<0.05),MDR1si1513组和MDR1si3071组的差别不大(P>0.05)。转染后72h,MDR1si326组蛋白质表达水平下降最明显。结论MDR1si326对人胃癌SGC7901/VCR细胞MDR1介导的耐药逆转效果最好,MDR1si2631次之,MDR1si3071较差,MDR1si1513最差。
Objective To screen the effective sequence of small interfering RNA (siRNA) targeting human multidrug resistance gene 1 (MDR1) in human gastric cancer cell line SGC7901 / VCR. Methods Four siRNAs targeting MDR1 (MDR1si326, MDR1si1513, MDR1si2631 and MDR1si3071) were designed and transfected into SGC7901 / VCR cells in vitro. The expression of MDR1 mRNA was detected by RT-PCR, the expression of P-glycoprotein (P gp) was detected by immunoblotting, the accumulation of adriamycin (ADR) in cells was detected by flow cytometry, the expression of P gp in cells was detected by MTT assay Sensitivity. Results All four siRNAs could reverse MDR1-mediated multidrug resistance in SGC7901 / VCR cells to varying degrees. At 48h after transfection, the mRNA expression levels of MDR1si326 group and MDR1si2631 group were 0.42 ± 0.07 and 0.49 ± 0.02, respectively, which were significantly lower than those of MDR1si1513 or MDR1si3071 group (P <0.05). The intracellular ADR accumulation in MDR1si326 group was the most significant 30.03, MDR1si2631 group, MDR1si3071 group, MDR1si1513 group at least (P <0.05); MDR1si2631 group had the highest relative reversal rate of ADR resistance (98.12 ± 0.26)%, MDR1si326 group (P <0.05) , There was no significant difference between MDR1si1513 group and MDR1si3071 group (P> 0.05). 72h after transfection, MDR1si326 group protein expression decreased the most obvious. Conclusion MDR1si326 has the best MDR1-mediated reversal effect on human gastric cancer cell line SGC7901 / VCR. MDR1si2631 is the second, MDR1si3071 is the worst, and MDR1si1513 is the worst.