三黄煎剂调节Aurora激酶A提高表柔比星对乳腺癌MDA-MB-231细胞药效的研究  

Sanhuang Decoction Regulating Aurora Kinase A and Promoting Epirubicine Chemosensitivity on MDA-MB-231 Cells

作　　者：卞卫和[1] 许岩磊[1] 陈曦琰 陈绪[1] 姚昶[1] 朱小纾[2] 张媞[1] 顾铭[1] 

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]西悉尼大学生物和健康学院,澳大利亚新南威尔士州彭里斯2750

出　　处：《辽宁中医杂志》2016年第4期798-803,I0003,共7页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81473676);江苏省高校优势学科建设项目Ⅱ期(012062003010);江苏省中医院高峰学术人才培养(y2014cr06)

摘　　要：目的:探讨三黄煎剂对表柔比星作用于MDA-MB-231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MDA-MB-231增殖的影响。RT-PCR法、Western Blot法检测三黄煎剂对MDA-MB-231细胞Aurora A、p53的mRNA及蛋白表达水平的影响。siRNA沉默MDA-MB-231细胞中Aurora A,并用CCK-8法检测对三黄煎剂作用于MDA-MB-231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MDA-MB-231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MDA-MB-231细胞凋亡相关蛋白及Aurora A表达的影响。结果:三黄煎剂对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)。三黄煎剂能够调节MDAMB-231细胞Aurora A、p53的mRNA及蛋白的表达。siRNA沉默Aurora A后,将三黄煎剂对MDA-MB-231细胞增殖抑制率下调了34.6%(51.5%到33.7%)。三黄煎剂与表柔比星联用能够增加表柔比星对MDA-MB-231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MDA-MB-231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。Objective： To explore the effect of Sanhuang Decoction on promoting Epirubicine chemosensitivity on MDA-MB-231 cells and regulating Aurora kinase A and discuss the underlying mechanism. Methods： The inhibition of MDA-MB-231 cells proliferation was determined by CCK-8 assay and the apoptosis detected by Annexin V-FITC / PI Staining. The knockdown of Aurora kinase A was made by Aurora A small interfering RNA（ siRNA）. The expression of Aurora A mRNA was examined by RT-PCR analysis. The expressions of apoptosis-related proteins and Aurora A were determined by Western blot analysis. Results： Sanhuang Decoction inhibited the proliferation of MDA-MB-231 cells in a dose-dependent manner（ P〈0. 05）. The effect of inhibition 48 hours after delivering to MDA-MB-231 cells was better than that after 24 hours（ P〈0. 05） and without statistical difference from that after 72 hours（ P〈0. 05）. Moreover,Sanhuang Decoction decreased the mRNA and protein level of Aurora A and increased those of p53 in a dose-dependent manner. The inhibition of MDA-MB-231 cells proliferation caused by Sanhuang Decoction dropped from 51. 5% to 33. 7% by application of Aurora A siRNA. Meanwhile,co-administration of Sanhuang Decoction and Epirubicine enhanced the effect of Epirubicine on inhibiting the proliferation,inducing the apoptosis and regulating apoptosis-related proteins and Aurora A. Conclusion： Combination of Sanhuang Decoction and Epirubicine increased the anticancer effect of Epirubicine,perhaps,through inhibiting Aurora kinase A.

关 键 词：三黄煎剂 Aurora激酶A MDA-MB-231细胞 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]