天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (3): 397-404.doi: 10.16333/j.1001-6880.2023.3.005

• 研究论文 • 上一篇    下一篇

山柰酚通过线粒体凋亡通路诱导三阴性乳腺癌细胞凋亡的机制研究

方雨潇,王淑美*   

  1. 重庆医科大学中医药学院 中医药防治代谢性疾病重庆市重点实验室,重庆 400016
  • 出版日期:2023-03-28 发布日期:2023-03-28
  • 基金资助:
    重庆市研究生科研创新项目(CYS21242);重庆市教委课题(22SKGH058)

Study on mechanism of kaempferol inducing apoptosis of triple negative breast cancer cells through mitochondrial apoptosis pathway

FANG Yu-xiao,WANG Shu-mei*   

  1. Chongqing Key Laboratory of TCM for Prevention and Cure of Metabolic Diseases,College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China
  • Online:2023-03-28 Published:2023-03-28

摘要:

本文旨在探讨山柰酚对三阴性乳腺癌细胞MDA-MB-231的抑制作用及其可能的分子机制。用不同浓度的山柰酚处理细胞,CCK-8法检测山柰酚对MDA-MB-231和人正常乳腺上皮细胞MCF-10A活力的影响,倒置显微镜观察各组细胞形态变化,克隆形成法检测细胞增殖,流式细胞仪检测细胞凋亡,JC-1染色法检测细胞线粒体膜电位的改变,Western blot检测B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞色素C(Cyt C)和细胞周期蛋白D1(CyclinD1)的蛋白表达,比色法检测半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3)以及半胱氨酸天冬氨酸蛋白水解酶-9(Caspase-9)的活性。结果显示,山柰酚体外可显著抑制MDA-MB-231细胞的增殖,诱导细胞凋亡,且对正常乳腺上皮细胞活力无影响,降低细胞线粒体膜电位,上调Bax、Cyt C的表达,下降Bcl-2和Cyclin D1的表达,增强Caspase-3、Caspase-9活性。结果表明,线粒体凋亡信号通路的激活是山柰酚诱导MDA-MB-231细胞凋亡的途径之一。

关键词: 柰酚, 三阴性乳腺癌, 线粒体凋亡通路, 细胞凋亡

Abstract:

The purpose of this study was to investigate the inhibitory effect of kaempferol on triple negative breast cancer cell MDA-MB-231 and its possible molecular mechanism.Cells were treated with kaempferol at different concentrations.CCK-8 method was used to detect the effect of kaempferol on the viability of MDA-MB-231 and human normal mammary epithelial cell MCF-10A.The morphological changes of cells in each group were observed by inverted microscope.Clone formation method was used to detect cell proliferation.Cell apoptosis was detected by flow cytometry.The changes of mitochondrial membrane potential were detected by JC-1 staining.Western blot was used to detect the protein expression of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cytochrome C(Cyt C) and Cyclin D1.Cysteinyl aspartate-specific protease 3(Caspase-3) and cysteinyl aspartate-specific protease 9 (Caspase-9) activities were detected by colorimetry.The results showed that kaempferol significantly inhibited the proliferation of MDA-MB-231 cells in vitro,induced apoptosis,and had no effect on the viability of normal mammary epithelial cells.It decreased the membrane potential of mitochondria,increased the expression of Bax and Cyt C,decreased the expression of Bcl-2 and Cyclin D1,and enhanced the activities of Caspase-3 and Caspase-9.It is suggested that the activation of mitochondrial apoptosis signal pathway is one of the pathways of kaempferol induced apoptosis in MDA-MB-231 cell.

Key words: kaempferol, triple negative breast cancer, mitochondrial apoptosis pathway, cell apoptosis

中图分类号:  R285