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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 investigate the effect and mechanism of sanhuang decoction on the effect of epirubicin on MDA-MB-231 cells. Methods: The CCK-8 method was used to detect the effect of Sanhuang decoction on the proliferation of breast cancer cells MDA-MB-231. The effects of Sanhuang decoction on mRNA and protein expression of Aurora A and p53 in MDA-MB-231 cells were detected by RT-PCR and Western Blot. siRNA was used to silence Aurora A in MDA-MB-231 cells and the effect of Sanhuang decoction on proliferation inhibition of MDA-MB-231 cells was detected by CCK-8 assay. CCK-8 assay and Annexin V-FITC / PI assay were used to detect the effects of Sanhuang Decoction combined with epirubicin on the proliferation inhibition rate and apoptosis rate of MDA-MB-231 cells. The effect of Sanhuang Decoction combined with epirubicin on apoptosis related protein and Aurora A expression in MDA-MB-231 cells was detected by Western Blot. Results: The inhibitory rate of Sanhuang decoction on MDA-MB-231 cells was increased in a gradient-dependent manner (P <0.05), and the curative effect of Sanhuang decoction for 48 hours was better than that of 24 h (P <0.05) Difference (P> 0.05). Sanhuang decoction can regulate the mRNA and protein expression of Aurora A and p53 in MDAMB-231 cells. Silencing Aurora A by siRNA decreased 34.6% (51.5% -33.7%) of Sanhuang decoction on the proliferation of MDA-MB-231 cells. Sanhuang decoction combined with epirubicin can increase the proliferation inhibitory rate and apoptosis rate of epirubicin on MDA-MB-231 cells, and regulate apoptosis-related protein c-PARP, c-Caspase 3 and Bcl-2 , Bax and Aurora A expression levels. Conclusion: Sanhuang decoction can increase the sensitivity of MDA-MB-231 cells to chemotherapeutic drug epirubicin, which may be related to the inhibition of Aurora kinase A by Sanhuang decoction.