Reversal effect of isotetrandrine, an isoquinoline alkaloid extracted from Caulis Mahoniae, on P-glycoprotein-mediated doxorubicin-resistance in human breast cancer (MCF-7/DOX) cells
- VernacularTitle:功劳木中异汉防己碱对P-糖蛋白介导的人乳腺癌细胞多药耐药性的逆转作用
- Author:
Tianxiao WANG
;
Xiaohong YANG
- Publication Type:Journal Article
- Keywords:
isotetrandrine;
P-glycoprotein;
multidrug resistance;
MCF-7/DOX cell
- From:
Acta Pharmaceutica Sinica
2008;43(5):461-466
- CountryChina
- Language:Chinese
-
Abstract:
This study investigated the reversal effect of isotetrandrine, an isoquinoline alkaloid extracted from Caulis mahoniae, on P-glyeoprotein-mediated multidrug resistance in human breast cancer doxorubicin-resistant (MCF-7/DOX) cells. RT-PCR assay and immunity histochemistry assay were used to determine the expression level of mdrl gene and P-gp in MCF-7/DOX cells to elucidate resistant character of MCF-7/DOX cells. The activity of isotetrandine to enhance doxorubicin cytotoxicity was tested using MTT (3-(4,5-dimethyhhiazol)-2,5-diphenyltetrazolium bromide) assay and was evaluated by the reversal fold (RF) values. Intracellular accumulation of doxorubicin was assessed by the determination of doxorubicin-assoeiated fluorescence intensity. Effect of isotetrandrine on the expression level of P-gp in MCF-7/DOX cells was then determined by immunity histochemistry assay. The ability of isotetrandrine to inhibit P-gp function was evaluated by detecting the accumulation and efflux of rhodamine123 ( Rh123 )with flow cytometry (FCM). Verapamil was employed as a comparative agent in whole experiment. The results indicated that MCF-7/DOX cells had phenotype of MDR and that the positive expression of P-gp MCF-7/DOX cells and reversal fold (RF) was significantly higher than that of verapamil (P < 0.05 ), butit hardly affected cytotoxicity of DOX in MCF-7 cells and the expression level of P-gp in MCF-7/DOXcells. The ability of isotetrandrine to inhibit P-gp function was reversible, because incubation of MCF-7/DOX cells with isotetrandrine caused a marked increase in uptake and a notable decrease in efflux of Rh123 and a marked increase of intracellular DOX concentrations. In conclusion, isotetrandrine exhibited potent effect on the reversal of P-gp-mediated MDR in vitro, suggesting that it might become a candidate of effective MDR reversing agent in cancer chemotherapy.
