【摘要】 目的通过重组腺病毒表达出mdr1基因的反义RNA,观察反义RNA对人肝癌基因工程细胞株HepG2/R多药耐药性的逆转效应。方法利用真核表达载体构建可以稳定表达mdr1基因和P糖蛋白的肝癌基因工程细胞株HepG2/R,通过腺病毒载体AdEasy系统生成重组腺病毒pAd Easy-GFP-ASmdr1,将此病毒感染HepG2/R,用RT-PCR检测mdr1 mRNA的表达强度,流式细胞仪检测细胞膜P-糖蛋白的表达和柔红霉素的蓄积率,HepG2/R细胞对阿霉素的耐药性用MTT法检测。结果HepG2/R细胞感染重组腺病毒后,RT-PCR检测表明mdr1 mRNA表达下降,流式细胞仪检测证实P-gp表达下降和柔红霉素的蓄积率增加,MTT法表明HepG2/R细胞对阿霉素的IC50从25μg/mL下降到3μg/mL。结论反义RNA通过抑制mdr1mRNA和P-gp的表达。更多还原

【Abstract】 【Objective】To investigate the reversal effect of multidrug resistance with antisense RNA of mdr1 gene delivered by recombinant adenoviruses in human engineering HCC cell line HepG2/R.【Methods】The recombinant adenoviruse was transfected into the engineering cell line HepG2/R.In order to investgate the reversal of the multidrug resistance phenotype, the expression of mdr1 mRNA was measured by RT-PCR, the production of P-glycoprotein and the accumulation of the daunorubicin(DNR) was determinated by flow cytometry.The sensitivitie of adriamycin(ADM) for HepG2/R cells was examined by MTT analysis.【Results】Compared with the parental HepG2 cells expressing low-level mdr1 mRNA and P-glycoprotein, engineering cell line HepG2/R was only stably expressing mdr1 gene and P-glycoprotion.The transfection of antisence RNA into HepG2/R cells resulted in decreases of mdr1 mRNA and P-glycoprotein levels.The sensitivity of transfected HepG2/R cells to ADM was reduced from 25μg/mL to 3μg/mL in IC50 level.The DNR accumulation was increased in transfected HepG2/R cells.ConclusionThis study demonstrates that mdr1 antisense RNA can increase the sensitivities of HepG2/R cells to anticancer drug by decreasing the expression of the mdr1 gene and inhibiting P-glycoprotein expression.更多还原