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[摘要]
目的 观察芒柄花素对不同亚型乳腺癌细胞周期相关基因和蛋白表达的影响。方法 乳腺癌细胞MCF-7、SK-BR-3、MDA-MB-231经0、40、80 μmol/L芒柄花素作用48 h后,分别采用Trizol和RIPA裂解细胞,收集细胞的总RNA和总蛋白,进一步采用实时定量PCR技术和Western blotting技术检测细胞周期调控因子cyclin D1、cyclin E、p21、p27的基因和蛋白表达情况。结果 3种乳腺癌细胞cyclin D1和cyclin E的基因及蛋白表达水平随着药物浓度的增加而降低,而p21和p27的基因和蛋白表达水平随药物浓度的增加而升高,与对照组比较,差异显著(P<0.05、0.01)。结论 芒柄花素可以通过调控cyclin D1、cyclin E、p21和p27基因和蛋白的表达,将乳腺癌细胞阻滞于G1期,从而抑制乳腺癌细胞的增殖。
[Key word]
[Abstract]
Objective To investigate the effect of formononetin on expression of cell cycle related genes and proteins in different subtypes of breast cancer cell lines. Methods Three subtypes of breast cancer cell lines, MCF-7, SK-BR-3 and MDA-MB-231, were treated with different concentrations of formononetin for 48 h. Total RNA and protein from breast cancer cells were extracted with Trizol and RIPA method. Then the gene and protein expression of cyclin D1, cyclin E, p21 and p27 were assayed by real-time PCR and Western blotting, respectively. Results After treated with formononetin, the levels of gene and protein expression of cyclin D1 and cyclin E were down-regulated in breast cancer cells, however those of p21 and p27 were up-regulated. Conclusion Formononetin could arrest breast cancer cell in G1 phase, through regulating the gene and protein expression of cyclin D1, cyclin E, p21 and p27.
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[基金项目]
福建省自然科学基金青年创新项目(2014D013);厦门市科技计划项目(3502Z20154052);厦门市中医后备人才项目