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目的:探讨MDR1siRNA对人舌癌耐药细胞株TCA8113多药耐药性的影响。方法:以人舌癌耐药细胞株TCA8113/DDP为靶细胞,将针对MDR1基因合成,并已鉴定和测序后的siRNA分子利用脂质体介导转染TCA8113/DDP细胞,采用G418筛选克隆细胞,RT-PCR检测MDR1基因mRNA表达,免疫组织化学法测定P-糖蛋白(P-gp)表达;MTT比色法检测TCA8113/DDP细胞对顺铂(DDP)、卡铂(CBP)、平阳霉素(PYM)的敏感性。结果:经转染成功的阳性克隆细胞TCA8113/DDP的MDR1mRNA和P-gp的表达均被抑制。TCA8113人舌癌耐药细胞顺铂(DDP)、卡铂(CBP)、平阳霉素(PYM)的IC50分别降至3.43μmol/L,1.72μmol/L和1.05μmol/L(P<0.01)。结论:稳定转染含编码MDR1siRNA的质粒载体能特异性地阻抑人舌癌多药耐药细胞TCA8113细胞MDR1基因的表达,逆转其对抗癌药物的耐受性。
Objective: To investigate the effect of MDR1 siRNA on the multidrug resistance of human tongue cancer cell line TCA8113. Methods: Human tongue cancer cell line TCA8113 / DDP was used as target cell. The siRNA against MDR1 gene was synthesized and transfected into TCA8113 / DDP cells by lipofectamine. The cloned cells were screened by G418. The mRNA expression of MDR1 was detected by RT-PCR and the expression of P-glycoprotein (P-gp) was detected by immunohistochemical method. The effect of TCA8113 / DDP on the expression of DDP, CBP, (PYM) sensitivity. Results: The expression of MDR1 mRNA and P-gp in transfected TCA8113 / DDP cells was inhibited. The IC50 of TCA8113 cells were decreased to 3.43μmol / L, 1.72μmol / L and 1.05μmol / L, respectively (P <0.01) with cisplatin (DDP), carboplatin (CBP) and pingyangmycin (PYM) CONCLUSION: Stable transfection of plasmid vector containing MDR1 siRNA can specifically inhibit the MDR1 gene expression in human tongue cancer multidrug resistance cell line TCA8113 and reverse its tolerance to anticancer drugs.