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目的探讨小干扰RNA(small interfering RNA,si RNA)对口腔鳞癌多药耐药基因mdr1(MDRI)及其表达产物P-糖蛋白(permeability-glyco-protein,P-gp)的干扰作用。方法体外构建针对MDR1的小干扰RNA,将其转染至人舌癌耐药细胞系Tca8113/DDP,采用RT-PCR法检测转染前后MDR1 m RNA的表达;采用免疫细胞化学技术比较转染前后P-gp的表达;采用MTT法检测转染前后肿瘤耐药细胞对顺铂的敏感性。结果 Tca8113/DDP细胞经MDR1-si RNA转染后48 h的转染率达最高,为71.3%;转染后mdr1 m RNA表达较对照组显著降低,降低率为68.32%,转染48 h后P-gp的表达较对照组明显降低;si RNA可显著提高Tca8113/DDP细胞对DDP的敏感性,逆转其耐药性,耐药倍数为2.05。结论 si RNA可以明显干扰口腔鳞癌MDR1及相应蛋白P-gp的表达。
Objective To investigate the interference of small interfering RNA (siRNA) on multidrug resistance gene mdr1 (MDRI) and its expression product P-glycoprotein (P-gp) in oral squamous cell carcinoma. Methods siRNA targeting MDR1 was constructed in vitro and transfected into human tongue cancer cell line Tca8113 / DDP. The expression of MDR1 m RNA before and after transfection was detected by RT-PCR. The expression of MDR1 mRNA was detected by immunocytochemistry before and after transfection P-gp expression; MTT assay before and after transfection of drug-resistant cells to cisplatin sensitivity. Results The transfection rate of Tca8113 / DDP cells transfected with MDR1-si RNA was the highest at 48 h, which was 71.3%. The expression of mdr1 m RNA in transfected Tca8113 / DDP cells was significantly lower than that in control group (68.32%). After 48 h of transfection, The expression of P-gp was significantly lower than that of the control group; si RNA could significantly increase the sensitivity of Tca8113 / DDP cells to DDP and reverse the drug resistance, with a multiple of 2.05. Conclusion si RNA can obviously interfere with the expression of MDR1 and P-gp in oral squamous cell carcinoma.