槐耳清膏体外逆转人乳腺癌细胞MCF-7耐三苯氧胺的研究

目的 探讨槐耳清膏体外逆转人乳腺癌细胞MCF-7耐三苯氧胺(TAM)的作用及其机制.方法 使用噻唑蓝(MTT)比色法测定不同药物处理对耐TAM的乳腺癌细胞株MCF-7/R的抑制率,流式细胞仪检测不同处理后MCF-7/R的凋亡情况和细胞周期变化.Western blot检测MCF-7/R不同处理后各组Phospho-P44/42MAPK(ERK1/2)及P44/42MAPK(ERK1/2)表达的差异.结果 MTT实验结果显示MCF-7/R的联合用药组抑制率(45.99±6.06)%和单药组抑制率(40.20±5.54)%分别与TAM对照组比较,差异有统计学意义(P＜0.05);流式细胞检测结果显示与TAM对照组比较,MCF-7/R的联合用药组和单药组G1期峰前出现显著的凋亡峰,且联合用药组的凋亡率(20.03%)与单药组的凋亡率(11.06%)与TAM对照组(2.15%)比较作用显著;Western blot检测蛋白表达经灰度分析显示,药物作用12 h后Phospho-P44/42MAPK(ERK1/2)蛋白在联合用药组的表达(0.3153±0.0179)和单药组的表达(0.5362±0.0030)分别与TAM对照组(0.9752±0.0034)比较均有下降,差异有统计学意义(P＜0.05),且联合组与单药组比较,差异有统计学意义(P＜0.05).结论 槐耳清膏具有体外逆转MCF-7/R细胞对TAM的耐药作用,逆转机制可能与下调MAPK通路磷酸化蛋白ERK1/2的表达水平相关,提示槐耳清膏是一种有意义的耐药逆转剂.

Experimental study on reversal of drug-resistance of human breast cancer cell line MCF-7 by Hua-ier fungi extract in vitro

Objective To investigate the effect and primary mechanism of Huaier fungi extract on reversal of tamoxifen-resistance of human breast cancer cell line MCF-7 in vitro. Methods Tamoxifen resistant cell line MCF-7/R cells were treated with the combination of Huaier fungi extract and Tamoxifen or separately, then the cell proliferation inhibition rates were determined by methyl thiazol tetrazolium (MTT)assay. The ratios of cells apoptosis and the change of cell cycle were also observed by flow cytometry. Protein expression of Phospho-P44/42MAPK(ERK1/2) and P44/42MAPK(ERK1/2) in each group were detected using Western blotting. Results It was demonstrated by MTT assay that the growth of MCF-7/R cells was inhibited in the combined group ( % ) (45.99 ± 6. 06) and single groups (40. 20 ± 5.54) % compared with control group (P ＜ 0. 05 ). It was revealed by flow cytometry that the MCF-7/R cell apoptosis rate were higher in the combined group (20. 03% ) and single groups ( 11.06% ) compared with control group (2. 15% ), and the combined group was the highest. Western blotting revealed that protein levels of Phospho-P44/42MAPK(ERK1/2) in combined group (0. 3153 ± 0. 0179) and single group (0. 5362 ±0. 0030) were inhibited significantly compared with control group (0. 9752 ± 0. 0034) at the 12-hour time point, and the difference is of significance in statistics science (P ＜ 0.05 ). Moreover, the difference between combined group and single group is of significance in statistics science ( P ＜ 0. 05 ). Conclusion Huaier fungi extract could reverse the tamoxifen-resistance of human breast cancer cell line MCF-7/R in vitro, and the reversal mechanism may related to the down-regulation of Phospho-P44/42 MAPK ( ERK1/2 )in MAPK pathway. It implied that Huaier fungi extract could be a significant reversal agent for clinic.