Effects of total flavone extract from Ampelopsis megalophylla mediated by autophagy inhibitor 3-MA on proliferation and apoptosis of human breast cancer cells
- VernacularTitle:基于自噬抑制剂3-MA介导的大叶蛇葡萄总黄酮提取物对人乳腺癌细胞增殖、凋亡的影响
- Author:
Shi-Yi XU
1
;
Si-Yu LIAO
;
Tian-Xu ZHANG
;
Xue ZOU
;
Chun GUI
;
Xiu-Qiao ZHANG
Author Information
- Keywords: total flavonoid extract of Ampelopsis mega-lophylla; inhibiting autophagy; breast cancer cells; MCF-7 cells; proliferation; apoptosis
- From: Chinese Pharmacological Bulletin 2024;40(6):1115-1123
- CountryChina
- Language:Chinese
- Abstract: Aim To explore the effect of total flavonoid extract(TFE)of Ampelopsis megalophylla on the pro-liferation and apoptosis of human breast cancer cells and its mechanism in autophagy inhibition.Methods For human cervical cancer cell Hela,human lung cancer cell A549,human liver cancer cell SMMC-7721,human breast cancer cell MCF-7,MDA-MB-231 and human normal liver cell L-02,MTT method was used to select sensitive cell lines.The inhibitory effect of TFE combined with autophagy inhibitor 3-methylade-nine(3-MA)on sensitive cell proliferation was detec-ted using MTT assay.The morphological changes of cells were observed using transmission electron micros-copy and Hoechst 33258 single staining method.The changes in cell apoptosis rate were detected using An-nexin V-FITC/PI dual staining method.The expression levels of apoptosis related proteins and pathway pro-teins(death receptor pathway,mitochondrial pathway,endoplasmic reticulum stress pathway)were detected uisng Western blot.The expression of the key protein Cyt-c in mitochondrial pathway was determiend by im-munofluorescence,and the autophagy agonist rapamy-cin was selected for reverse validation.Results TFE could inhibit the proliferation of human breast cancer cells in a concentration-dependent manner,and MCF-7 cells were sensitive cell lines.Compared with the TFE group,the TFE+3-MA group significantly increased the inhibition rate of MCF-7 cells at 24,48,and 72 h(P＜0.01).The number of cells decreased,the gap increased,the number of apoptotic bodies increased,and the apoptosis rate increased(P＜0.01).The ex-pression levels of Bax/Bcl-2(P＜0.01),cleaved-caspase3(P＜0.01),Cyt-c(P＜0.05),FADD,and cleaved-caspase-12 all increased,and the expres-sion of apoptotic protein Cyt-c in nucleus increased.The fluorescence of the TFE+RA group decreased,re-versing the mitochondrial pathway apoptosis induced by TFE.Conclusions TFE can significantly inhibit the proliferation of human breast cancer cells.When inhib-iting autophagy,it may promote the apoptosis of MCF-7 cells through the mitochondrial pathway,and activa-ting autophagy can reverse apoptosis.
