黄芪对肝癌耐药细胞株Bel/Fu化疗敏感性的影响

Effect of Astragulus Injection on Chemosensitivity of Drug-resistant Hepatic Carcinoma Bel/Fu Cell Line

目的:评估黄芪对肝癌耐药细胞化疗敏感性的影响,探讨黄芪对血红素加氧酶调节作用的可能机制。方法:黄芪注射液作用于Bel/Fu肝癌化疗耐药细胞株,分别应用MTT、免疫细胞化学、流式细胞术、RT-PCR检测细胞株对化疗药物敏感性、P-糖蛋白表达量、P-糖蛋白功能、血红素加氧酶(HO-1)核酸水平表达量。结果:黄芪注射液作用于Bel/Fu细胞诱导24h后,化疗药物半数抑制浓度显著降低;P-糖蛋白阳性表达明显减少,黄芪组罗丹明荧光曲线明显右移,细胞化疗药物外排功能降低;HO-1 mRNA表达显著减少。结论:黄芪注射液可增强肝癌耐药细胞株Bel/Fu的化疗敏感性,下调P-糖蛋白表达,降低P-糖蛋白药物外排功能,实现这种作用的同时伴随有HO-1 mRNA表达的减少。

Objective To study the effect and relevant mechanism of Astragulus injection on chemosensifivity of drug -resistant hepatic carcinoma Bel/Fu cell line. Methods The drug sensitivity test of Bel/Fu was carried out with /WIT assay. Thc distribution of P-glucoprotein（P-GP） in drug-resistant cells were determined by immunohistocbemical staining. Rhodamine 123 retention fluorescent intensity measured by flow cytometry was used to detect the function of P - GP. HO-1mRNA level was determined by RT-PCR technique. Results After Bel/Fu cells being dealing with Astragulus injection for 24 h, there were significant differences in 50% inhibiting concentration （ICs0）of chemical drugs between control groups and contrast group（ P 〈 O. O1 ）. IC50 of control group decreased obviously. P-GP expression were significantly lower in the cells treated with 8 uL/mL Astragulus injection than those of contrast group （Z = - 2. 363, P = O. 018）. The curve of Rhodamine 123 retention fluorescent intensity in the control cells removed rightward from thc place of contrast group indicating that the efflux pump function of P-GP was weakened. Meanwhile, the mRNA cxpression of HO-1 in the cells of control group were observably lower than those of contrast group（O. 316 ＋ O. 08 vs O. 624 ＋ O. 173, P 〈 O. 01）. Conclusion . Astragulus injection can enhance the chemosensitivity of Bel/Fu cell line, that may be related to decreasing protein expression and efllux pump function of P-GP concomitant with the decrease of HO-1mRNA lcvcl.