三黄煎剂调节Aurora激酶A促进乳腺癌细胞凋亡的实验研究
San Huang Decoction Promotes Apoptosis of Breast Cancer Cells Through Regulating Aurora Kinase A
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摘要: 目的 探讨三黄煎剂对乳腺癌MCF-7、MDA-MB-231细胞凋亡的影响及Aurora激酶A(Aurora A)蛋白表达及功能的影响,并探讨其内在机制。方法 采用CCK-8法检测三黄煎剂对乳腺癌细胞MCF-7、MDA-MB-231增殖的影响。AnnexinV-FITC/PI法检测MCF-7、MDA-MB-231细胞凋亡率。q-PCR法检测MCF-7、MDA-MB-231细胞Aurora A、p53的mRNA表达水平。Western Blot法检测MCF-7、MDA-MB-231细胞凋亡相关蛋白的表达及Aurora A蛋白的表达。结果 三黄煎剂对MCF-7、MDA-MB-231细胞增殖抑制率呈浓度梯度依赖增长(P<0.05),给药48 h疗效好于24 h(P<0.05),与给药72 h无统计学差异(P>0.05)。三黄煎剂能够诱导MCF-7、MDA-MB-231细胞凋亡,并上调c-PARP、c-Caspase 3、Bax蛋白的表达,下调Bcl-2蛋白的表达,呈浓度梯度依赖。三黄煎剂能够下调Aurora A蛋白及mRNA的表达、上调p53蛋白及mRNA的表达。结论 三黄煎剂能够通过下调Aurora A蛋白及mRNA的表达,抑制Aurora A的生物活性,抑制MCF-7、MDA-MB-231细胞增殖,诱导其凋亡。Abstract: OBJECTIVE To explore the effect of San Huang decoction on the apoptosis of breast cancer cells and the effect on the mRNA and protein expression and function of Aurora kinase A and discuss the underlying mechanism of San Huang induced apoptosis. METHODS The inhibition of breast cancer cells proliferation was determined by CCK-8 assay. The apoptosis of breast cancer cells was detected by AnnexinV-FITC/PI Staining. The expression of mRNA of Aurora A was examined by q-PCR analysis. The expression of apoptosis-related proteins and Aurora A were determined by Western Blot analysis. RESULTS San Huang decoction inhibited the proliferation of breast cancer cells in a does-dependent manner(P<0.05). The effect of inhibition caused by San Huang decoction 48 hours after delivering to breast cancer cells was better than 24 hours(P<0.05) , although similar as 72 hours(P>0.05). San Huang decoction was also found to induce apoptosis in both MCF-7 and MDA-MB-231 cell lines in a dose-dependent manner. Consistent with cellular results, San Huang decoction treatment significantly increased the apoptosis-related protein level of cleaved-PARP(c-PARP), cleaved-Caspase 3(c-Caspase 3) and Bax, down-regulated Bcl-2 in a does-dependent manner. Meanwhile, San Huang decoction decreased the mRNA and protein level of Aurora A and increased those of p53 in a does-dependent manner. CONCLUSION San Huang decoction at the first time was able to promote the apoptosis of breast cancer cells via inducing the suppression of Aurora A.