不同浓度中药柴胡提取物对人肝癌细胞内Ca^(2+)和p53表达的影响

Effects of the Extracts from BCDC on Intracelluar Calcium and p53 of Human Hepatoma Cells

目的:测定不同浓度中药柴胡(Bup leurun Ch inese DC,BCDC)提取物对BEL-7402细胞内游离钙离子浓度([Ca2+]i)和p53表达的影响,以探索柴胡逆转BEL-7402细胞多药耐药的机制。方法:用Fura-2/AM作为细胞内钙离子的荧光指示剂,用双波长荧光分光光度计测定不同浓度柴胡提取物作用下BEL-7402细胞内[Ca2+]i及其剂量-效应关系;用免疫组化法检测不同条件下BEL-7402细胞的p53表达。结果:柴胡可使人肝癌细胞BEL-7402细胞内游离钙离子浓度下降(P<0.001),但不具有剂量-效应关系;BEL-7402细胞呈p53天然突变。结论:柴胡可使人肝癌细胞BEL-7402细胞内游离钙离子浓度下降,为一种钙离子通道阻滞剂(calc ium channel b locker,CCB),提示钙离子通道阻滞作用为柴胡逆转BEL-7402细胞多药耐药的机制之一;BEL-7402细胞呈p53天然突变也可直接或间接影响肿瘤多药耐药性的表达。

Objective： To measure the effects of the extracts from BCDC on intracellular free calcium and the expression of p53 of human hepatoma BEL-7402 cells, in order to explore the mechanism of BCDC reversing the multidrug resistance of BEL-7402 ceils of human hepatoma. Methods： Double wavelength spectrofluorometer with Fura-2/AM as the fluorescence indicator for intracellular free calcium ions （ [ Ca^2＋ ] i） was used to measure the changes of [ Ca^2＋ ] i in BEL-7402 cells of human hepatoma affected by BCDC and the expression of p53 was examined by immunochemical assay. Results： BCDC can reduce intracellular free calcium of human hepatoma BEL-7402 cells （P 〈 0. 001 ）. Immunochemical assay showed that BCDC had no effect on p53 gene expression. Conclusion： BCDC is one of calcium channel blocker and is able to reduce intracellular free calcium of human hepatoma BEL-7402 ceils ,which indicats that the effect of calcium channel blocker may be one of the mechanism of BCDC reversing the mutidrug resistance of BEL-7402 ceils of human hepatoma. And the p53 mutation of BEL-7402 ceils can affect the expression of MDR1.