小柴胡汤逆转人肝癌细胞BEL-7402／5-FU 的多药耐药作用

目的:研究小柴胡汤对BEL-7402/5-FU细胞多药耐药的逆转作用,并初步探讨其作用机制。方法:采用四甲基偶氮唑蓝(MTT)法检测不同化疗药物以及小柴胡汤对BEL-7402/5-FU细胞的半数抑制浓度(IC50),计算耐药指数和小柴胡汤的逆转倍数;流式细胞仪检测细胞内阿霉素的荧光强度;RT-PCR检测mdr1基因表达;western blot检测p-gp蛋白表达。结果:BEL-7402/5-FU耐药株对5-FU和阿霉素顺铂均具有耐药性,耐药指数分别是6 501.99和31.73;0.5 mg/ml小柴胡汤和5-FU、阿霉素联合应用,其逆转倍数分别是1.66、1.50;1.0 mg/ml小柴胡汤和5-FU、阿霉素联合应用,其逆转倍数分别是3.44、5.58;小柴胡汤增强阿霉素的荧光强度;小柴胡汤降低mdr1基因表达和p-gp蛋白表达,各个浓度均具有统计学意义。结论:小柴胡汤可以抑制mdr1基因编码蛋白产物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。

Xiaochaihu Tang reverses multidrug resistance in resistant human hepatocellular carcinoma cell line BEL-7402/5-FU

Objective: To determine the reversal effect of Xiaochaihu tang (XCHT) on the resistant human hepatocellular carcinoma cell line BEL-7402/5-FU and explore its mechanism. Methods: The IC50 of BEL-7402/5-FU cells was examined by MTT and calculated the resistance index (RI) and the reversal folds (RF) of XCHT. Intracellular Adriamycin (ADM) concentration was determined by flow cytometry. RT-PCR and western blot was used to detected the expression of mdr1 and p-gp respectively. Results: BEL-7402/5-FU showed resistance to variety of chemotherapeutic drugs, the RI of 5-FU or ADM was 6 501.99 or 31.73.XCHT (0.5 mg/ml or 1.0 mg/ml) combined with 5-FU and ADM, the reversal fold was 1.66 and 1.50 or 3.44 and 5.58. XCHT increased the intracellular concentration of ADM. XCHT down-regulated the mdr1 and p-pg expression, and both had significant difference. Conclusion: XCHT can inhibit the expression of p-gp coded by mdr1 gene, and increase the intracellular concentration of chemotherapeutic drugs, so as to reverse the multidrug resistance of HCC cells.