索拉非尼逆转肝癌细胞多药耐药的实验研究

Sorafenib reverses multidrug resistance of hepatoma cells in vitro

目的探讨索拉非尼体外对人肝癌细胞(BEL-7402/FU)多药耐药性的逆转作用及可能机制。方法以MTT比色法测定索拉非尼对肝癌细胞的剂量效应曲线,并以流式细胞仪检测索拉非尼对肝癌细胞内罗丹明123(Rho123)浓度的影响,根据上述实验结果选取合适的索拉非尼实验剂量。以MTT法测定索拉非尼对抗癌药物细胞毒性的影响,用流式细胞仪检测细胞膜转运蛋白(P-gp)的表达,RT-PCR法检测mdr1基因的表达。结果索拉非尼浓度在4μmol/L时逆转效率较高且毒副作用较小,4μmol/L的索拉非尼能部分逆转BEL-7402/FU细胞的耐药性,对ADM、5-FU、GEM、DDP的逆转倍数分别为2.98、7.16、1.99、10.08倍,并可部分下调BEL-7402/FU细胞P-gp的表达,使mdr1基因表达与对照组相比下降了27.3%。结论索拉非尼具有体外逆转人肝癌细胞多药耐药性的作用,可能与下调mdr1基因表达、增加细胞内化疗药物的蓄积有关。

Objective To explore the role of sorafenib in reversing multidrug resistance （MDR） in hepatoma BEL-7402/FU cells and its possible mechanisms. Methods MTT colorimetric assay was used to obtain the dose-response curve of sorafenib in BEL-7402/FU cells, and flow cytometry performed to assess the effect of sorafenib on Rho123 concentration in the cells. The optimal dose of sorafenib for cell treatment was determined according to the results of MTT assay and flow cytometry. MTT assay was employed to evaluate the effect of sorfenib on the cytotoxicity of the antitumor drugs, flow cytometry performed to determine the expression of cell membrane transport protein （P-gp）, and RT-PCR used to detect mdrl gene expression in the ceils treated with sorafenib at the optimal dose. Results Sorafenib at the concentration of 4 μmol/L efficiently reversed the MDR of the cells with minimal side effects. At the concentration of 4 μmol/L, sorafenib partially reversed the drug resistance of BEL-7402/FU cells to ADM, 5-FU, GEM and DDP, with reversal indexes of 2.98, 7.16, 1.99 and 10.08, respectively. Treatment of the cells with 4 μmol/L sorafenib also partially down-regulated P-gp expression in BEL-7402/FU cells, and caused a reduction of mdrl gene expression by 27.3% in comparison with the control cells. Conclusion Sorafenib can reverse MDR in human hepatoma cells probably in association with down-regulation of mdrl gene expression and increased accumulation of the chemotherapeutic agents in the cells.