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[目的]探讨应用RNAi技术抑制肝癌耐药细胞MDR1基因表达的抑制作用以及对P-糖蛋白(P-gp)表达和功能的抑制作用。[方法]根据MDR1基因序列,设计并体外转录合成2条siRNA(smallinterferingRNA),用脂质体转染将其导入细胞内。应用MTT检测转染后癌细胞生长增殖情况,用MTT法测定细胞对化疗药物阿霉素(ADM)的敏感性,流式细胞仪检测细胞膜表面P-糖蛋白(P-gp)表达和细胞内罗丹明(Rhdaming123,Rh123)的潴留。[结果]转染sh-MDR1-1和sh-MDR1-2可显著抑制两株耐药细胞MDR1mRNA和P-gp的表达,细胞内的Rh123稳态积累量均明显增高;第1条序列更能有效的抑制MDR1基因表达。[结论]sh-MDR1-1特异性siRNA可更强的抑制肝癌耐药细胞MDR1基因表达。
[Objective] To investigate the inhibitory effect of RNAi on the MDR1 gene expression and the inhibitory effect on the expression and function of P-glycoprotein (P-gp) in hepatocellular carcinoma cell line. [Method] According to the MDR1 gene sequence, two small interfering RNAs were designed and synthesized by in vitro transcription and transfected into cells by lipofectamine. The growth and proliferation of cancer cells were detected by MTT assay. The cell chemosensitivity to doxorubicin (ADM) was determined by MTT assay. The expression of P-glycoprotein (P-gp) Rhodamine (Rh123) retention. [Results] Transfection of sh-MDR1-1 and sh-MDR1-2 significantly inhibited the expression of MDR1 mRNA and P-gp in both drug-resistant cells and the steady-state accumulation of Rh123 in cells was significantly increased. Effectively inhibit MDR1 gene expression. [Conclusion] sh-MDR1-1 specific siRNA can inhibit the MDR1 gene expression in multidrug resistant hepatocellular carcinoma cells more strongly.