索拉非尼对耐药肝癌细胞株BEL-7402/FU逆转作用的研究

Reversion of drug-resistant hepatocellular carcinoma cell line BEL-7402/FU by sorafenib

目的:研究索拉非尼对肝癌耐药细胞系BEL-7402/FU多药耐药性的逆转作用及可能机制。方法:采用流式细胞术检测罗丹明123(Rho123)的表达,免疫组化法观察细胞膜上P-gp的表达,荧光定量PCR法检测多药耐药基因mdr1 mRNA水平的变化,Western blot法检测mdr1基因蛋白产物P-gp表达水平的变化。结果:经4μmol/L索拉非尼处理后,增加了BEL-7402/FU对Rho123的蓄积并减缓其外排作用,使细胞表面P-gp的表达明显下降,mdr1 mRNA较用药前下降了35.4%,并可明显抑制mdr1基因蛋白产物P-gp的表达水平,比BEL-7402/FU细胞减少14.3%。结论:索拉非尼可以抑制mdr1基因蛋白产物P-gp的表达,增加细胞内化疗药物的浓度,从而逆转了肝癌细胞的多药耐药性。

AIM： To investigate the reversion of multi- drug resistance of drug-resistant hepatocellular carcinoma （HCC） cell line BEL-7402/FU by sorafenib and the possible mechanisms. METHODS： Flow cytometry was applied to detect the expression of rhodamine 123 （ Rho123）, immunehistochemistry was applied to observe the expression of P- glycoprotein （P-gp） on the cell membrane, the fluorescence quantitative PCR assay was applied to determine the changes of the multidrug-resistant gene mdr1 mRNA level and the Western blot assay was applied to test the changes of the expression level of the mdr1 gene product P-gp. RESULTS： After 4 μmol/L treatment by sorafenib, Rho123 accumulation was increased by BEL-7402/FU and exocytosis was slowed down, which evidently reduced the expression of cell surface P-gp; the mdrl mRNA level was reduced by 35.4% compared to its level prior to drug administration; the expression level of the mdrl gene product P-gp was significantly suppressed, 14.3% fewer than that of BEL-7402/FU cells. CONCLUSION： Sorafenib can inhibit the expression of the mdrl gene product P-gp and increase the intracellular concentration of chemotherapeutic drugs, so as to reverse the multidrug resistance of HCC cells.