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。
[ Objective ] To investigate the reversal effect of multidrug resistance with antisense RNA of mdrl 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 mdrl mRNA was measured by RT-PCR, the production of P-glyco- protein and the accumulation of the daunorubicin （DNR） was determinated by flow eytometry. The sensitivitie of adriamyein （ADM） for HepG2/R cells was examined by MTT analysis. [ Results ] Compared with the parental HepG2 cells expressing low-level mdrl mRNA and P-glycoprotein, engineering cell line HepG2/R was only stably expressing mdrl gene and P-glyeoprotion. The transfeetion of antisence RNA into HepG2/R cells resulted in decreases of mdrl 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. [Conclusion] This study demonstrates that mdrl antisense RNA can increase the sensitivities of HepG2/R cells to anticaneer drug by decreasing the expression of the mdrl gene and inhibiting P-glyeoprotein expression.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2009年第19期2907-2910,2914,共5页
China Journal of Modern Medicine
基金
泸州医学院和四川省科技厅科研资助项目(No:05JY029-146)
关键词
多药耐药性
多药耐药基因
肝癌细胞株
工程细胞株
mulfidrug resistance
muhidrug resistance gene
hepatoeellular carcinoma cell line
engineering cell line
