摘要
目的观察薯蓣皂苷元对乳腺癌细胞增殖与迁移的抑制作用,探讨其作用机制与NF-κB信号通路的关系。方法采用CCK-8法检测不同浓度的薯蓣皂苷元作用乳腺癌细胞株及正常细胞株24 h后的生长抑制率。倒置相差显微镜下观察薯蓣皂苷元作用MDA-MB-231细胞24 h后,细胞形态的变化。细胞划痕法观察薯蓣皂苷元对MDA-MB-231细胞迁移的影响。RT-PCR检测凋亡相关基因Bcl-2 mRNA表达情况。Western Blot法检测薯蓣皂苷元对MDA-MB-231细胞凋亡相关蛋白Bax、Bcl-2的影响,以及对NF-κB信号通路相关蛋白P65的影响。结果不同浓度薯蓣皂苷元对两种人类三阴性乳腺癌细胞株和小鼠E0771细胞均有显著的抑制效应,但对End1/E6E7细胞影响较小,只有在极高浓度作用下才产生抑制效应。薯蓣皂苷元作用MDA-MB-231细胞后,细胞的形态发生改变,细胞边缘回缩,失去正常上皮生长样形态。薯蓣皂苷元可显著抑制MDA-MB-231细胞的迁移,并呈浓度和时间依赖性。薯蓣皂苷元作用MDA-MB-231细胞24 h后,Bcl-2 mRNA表达量低于对照组。Bcl-2、P65蛋白表达量低于空白组,Bax蛋白表达量高于对照组。结论薯蓣皂苷元对乳腺癌细胞的抑制作用明显,其作用机制可能与抑制NF-κB信号通路的激活有关。
Objective To observe the inhibitory effect of diosgenin on the proliferation and migration of breast cancer cells and explore the relationship between its mechanism of action and the NF-κB signaling pathway. Methods CCK-8 method was used to detect the growth inhibition rate of diosgenin-treated breast cancer cell lines and normal cell lines after different concentrations for 24 hours. Under inverted phase contrast microscope, the morphological changes of MDA-MB-231 cells treated with diosgenin for 24 hours were observed. RT-PCR was used to detect the expression of Bcl-2 mRNA. Western Blot was used to detect the effect of diosgenin on apoptosis-related proteins Bax and Bcl-2 in MDA-MB-231 cells and the effect on NF-κB signaling pathway-related protein P65. Cell scratch method was used to observe the effect of diosgenin on the migration of MDA-MB-231 cells. Results Diosgenin at different concentrations had an inhibitory effect on breast cancer cell lines, especially in E0771 cells. At the same time, diosgenin had a small effect on End1/E6 E7 cells, and the inhibitory effect was produced only at very high concentrations. After diosgenin acting on MDA-MB-231 cells, the morphology of the cells changed, the cell edges shrank and the normal epithelial growth-like morphology was lost. The expressions of Bcl-2 mRNA in MDA-MB-231 cells treated with diosgenin for 24 hours was lower than that in the control group. The expressions of Bcl-2 and P65 protein were lower than those of the blank group, and the expression of Bax protein was higher than that of the control group. Diosgenin significantly inhibited the migration of MDA-MB-231 cells in a concentration and time-dependent manner. Conclusion Diosgenin has obvious inhibitory effect on breast cancer cells, and its mechanism may be related to inhibiting the activation of NF-κB signaling pathway.
作者
孙晓东
朱方怡
谢丽霞
徐红霞
葛京京
桑明
SUN Xiaodong;ZHU Fangyi;XIE Lixia;XU Hongxia;GE Jingjing;SANG Ming(Xiangyang NO.1 People's Hospital,Hubei Key Laboratory of Wudang Local Chinese Medicine Research,Hubei University of Medicine,Xiangyang 442000,Hubei,China)
出处
《中华中医药学刊》
CAS
北大核心
2021年第6期112-116,I0026,共6页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(81903005)
湖北省自然科学基金(2017CFB674,2018CFB701)
湖北省对外科技合作项目(2019AHB068)
湖北省卫生与健康委员会科研项目(ZY2019F028)
武当特色中药研究湖北省重点实验室(湖北医药学院)开放课题项目(WDCM2018009)。
