黄芩素联合U0126诱导人乳腺癌细胞株MCF-7凋亡的分子机制研究

发布时间：2018-04-02 07:03

本文选题：黄芩素　切入点：U　出处：《中国现代医学杂志》2017年10期

【摘要】：目的探讨黄芩素和U0126诱导人乳腺癌细胞株MCF-7凋亡的分子机制。方法单独及联合使用20μmol黄芩素、10μmol U0126处理MCF-7细胞24 h;光学显微镜观察细胞数量变化,流式细胞术检测MCF-7细胞周期变化,CCK8法检测细胞增殖变化,TUNEL法和流式细胞术检测细胞凋亡,实时聚合酶链反应(Real Time PCR)和Western blot检测凋亡相关因子m RNA和蛋白的表达水平。结果与黄芩素单独刺激MCF-7细胞相比,黄芩素联合U0126刺激MCF-7细胞时,S期细胞比例降低更明显;MCF-7细胞经不同浓度黄芩素或U0126处理后,增殖抑制,且具有浓度依赖性(P0.05);与黄芩素单独处理MCF-7细胞相比,黄芩素与U0126联合处理时,细胞凋亡更加显著(P0.05),早期凋亡和晚期凋亡均增多(P0.05);与黄芩素或U0126单独处理MCF-7细胞相比,黄芩素和U0126联合处理MCF-7细胞时,凋亡抑制因子Bcl-2的m RNA水平降低(P0.05),凋亡促进因子Bax的m RNA水平增高(P0.05),ERK1/2、GSK-3β和P38的磷酸化水平降低(P0.05),凋亡促进因子Bax表达量增高(P0.05),凋亡抑制因子Bcl-2表达量降低(P0.05)。结论黄芩素或U0126通过调控Bcl-2/Bax的表达水平以及ERK1/2、GSK-3β和P38的磷酸化水平抑制MCF-7细胞增殖,促进细胞凋亡,且具有协同效应。因此,黄芩素和U0126可联合用于临床治疗乳腺癌,具有重要临床意义。
[Abstract]:Objective to investigate the molecular mechanism of baicalin and U0126 inducing apoptosis in human breast cancer cell line MCF-7. Methods MCF-7 cells were treated with 20 渭 mol baicalin 10 渭 mol U0126 for 24 h, and the number of MCF-7 cells was observed by optical microscope. MCF-7 cell cycle changes were detected by flow cytometry. Cell proliferation was detected by CCK8 method and apoptosis was detected by Tunel and flow cytometry. Real-time polymerase chain reaction (Time) and Western blot were used to detect the expression of apoptosis-related factor m RNA and protein. Results compared with baicalin alone, the expression of m RNA and protein was detected in MCF-7 cells. The ratio of S phase cells in MCF-7 cells stimulated by baicalin combined with U0126 was significantly decreased after treated with different concentrations of baicalin or U0126, the proliferation of MCF-7 cells was inhibited in a concentration-dependent manner, compared with that of MCF-7 cells treated with baicalin alone. When combined with baicalin and U0126, the apoptosis of MCF-7 cells was significantly higher than that of baicalein and U0126, and increased in both early and late stages of apoptosis. Compared with baicalin or U0126 alone, baicalein and U0126 treated MCF-7 cells. The level of m RNA of apoptosis inhibitor Bcl-2 decreased P0.05, the level of m RNA of Bax increased and the phosphorylation of P0.05 / ERK1 / 2 + GSK-3 尾 and P38 decreased, the expression of Bax increased and Bcl-2 decreased P0.055.Conclusion the expression of Bcl-2 is lower than that of Bax. Baicalin or U0126 inhibited the proliferation of MCF-7 cells by regulating the expression of Bcl-2/Bax and phosphorylation of ERK1 / 2, GSK-3 尾 and P38. The combination of baicalin and U0126 can be used in clinical treatment of breast cancer and has important clinical significance.
【作者单位】：四川省泸州市人民医院乳腺外科;西南医科大学附属中医医院妇产科;西南医科大学附属医院医学实验中心;西南医科大学附属医院妇产科;西南医科大学医学信息与工程学院;
【基金】：四川省科技厅项目(14JC01353-LH67) 四川省泸州市科技局项目(2014-S-44)
【分类号】：R737.9

【相似文献】