三黄煎剂抑制Aurora激酶A增强表柔比星对乳腺癌MCF-7细胞药效的研究

Study on Improvement of Chemosensitivity of MCF-7 Cells to Epirubicin and Inhibition of Aurora Kinase A in Treatment of Breast Cancer by San-Huang Decoction

目的:探讨三黄煎剂对表柔比星作用于MCF-7细胞药效的影响及相关机制。方法:采用CCK-8法检测三黄煎剂对乳腺癌细胞MCF-7增殖的影响。RT-PCR法、Western Blot法检测三黄煎剂对MCF-7细胞Aurora A、p53的m RNA及蛋白表达水平的影响。si RNA沉默MCF-7细胞中Aurora A,并用CCK-8法检测三黄煎剂对MCF-7细胞增殖抑制的影响。CCK-8法、Annexin V-FITC/PI法检测三黄煎剂联合表柔比星对MCF-7细胞增殖抑制率、凋亡率的影响。Western Blot法检测三黄煎剂联合表柔比星对MCF-7细胞凋亡相关蛋白表达的影响。结果:三黄煎剂对MCF-7细胞增殖抑制率呈浓度梯度依赖增长(P<0.05),给药48 h疗效优于24 h(P<0.05),与给药72 h无统计学差异(P>0.05)。三黄煎剂能够调节MCF-7细胞Aurora A、p53蛋白及m RNA的表达。si RNA沉默Aurora A将三黄煎剂对MCF-7细胞的增殖抑制率下调了50.0%(49.2%到24.8%)。三黄煎剂与表柔比星联用能够增加表柔比星对MCF-7细胞的增殖抑制率及凋亡率,调节凋亡相关蛋白c-PARP、c-Caspase 3、Bcl-2、Bax及Aurora A的表达水平。结论:三黄煎剂能够增加MCF-7细胞对化疗药物表柔比星的敏感性,可能与三黄煎剂对Aurora激酶A的抑制有关。

This article was aimed to explore the effect of San-Huang（SH） decoction on improving chemosensitivity of MCF-7 cells to epirubicin and inhibition of Aurora kinase A, in order to discuss its underlying mechanism. The inhibition of MCF-7 cells proliferation on breast cancer by SH decoction was determined by CCK-8 assay. RT-PCR and western blot were used to detect the Aurora A, p53 m RNA and protein expression level of MCF-7 cells by SH decoction. The si RNA silenced Aurora A of MCF-7 cells. CCK-8 assay was used to detect the inhibition of MCF-7 cells proliferation. CCK-8 assay and Annexin V-FITC/PI staining were used to detect the inhibition rate and apoptosis rate of MCF-7 cells treated by the combination of SH decoction and epirubicin. Western blot analysis was used to detect the expression of apoptosis-related proteins. The results showed that SH decoction inhibited the proliferation of MCF-7 cells in a dose-dependent manner（P〈0.05）. The effect of 48 h medication was better than 24 h（P〈0.05）. There was no statistical difference with medication for 72 h（P〉0.05）. SH decoction can regulate the Aurora A, p53 protein and m RNA expression of MCF-7 cells. si RNA silenced Aurora A, which downregulated the inhibition rate of MCF-7 cells by SH decoction for 50.0%（from 49.2% to 24.8%）. The combination of SH decoction and epirubicin enhanced the effect of epirubicin on inhibiting the proliferation rate and apoptosis rate of MCF-7 cell, regulated the expression levels of apoptosis-related protein such as c-PARP, c-Caspase 3, Bcl-2, Bax, as well as the protein level of Aurora A. It was concluded that SH decoction can increase the chemosensitivity of MCF-7 cells to epirubicin, which may be related to the inhibition of Aurora Kinase A by SH decoction.