Mechanism of Polyphyllin Ⅱ in Induction of Ferroptosis in Hepatocellular Carcinoma HepG2 Cells
10.13422/j.cnki.syfjx.20240629
- VernacularTitle:重楼皂苷Ⅱ诱导肝癌HepG2细胞铁死亡作用机制
- Author:
Huizhong ZHANG
1
;
Jian NI
1
;
Hulinyue PENG
1
;
Yibo ZHANG
1
;
Xiaohan XU
1
;
Shiman LI
1
;
Yidan RUAN
1
;
Yongqiang ZHANG
1
;
Pingzhi ZHANG
1
;
Aina YAO
1
;
Ying WANG
2
;
Xiaoxu DONG
1
Author Information
1. School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing 102488， China
2. National Institutes for Food and Drug Control，Beijing 100050，China
- Publication Type:Journal Article
- Keywords:
hepatocarcinoma carcinoma;
polyphyllin Ⅱ;
ferroptosis;
p53;
mechanism
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(17):105-112
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the induction of ferroptosis by polyphyllin Ⅱ （PPⅡ） in hepatocellular carcinoma HepG2 cells and its underlying mechanism. MethodThe effect of PPⅡ （0， 1.5， 3.0， 4.5， 6.0， 9.0， 18.0 mg·L-1） on the in vitro proliferation of HepG2 cells was assessed using the methyl thiazolyl tetrazolium （MTT） assay. Colony formation ability of HepG2 cells was evaluated through a colony formation assay. Cell migration ability was assessed via a scratch assay. Lactate dehydrogenase （LDH） content in HepG2 cells was measured using a kit. Reactive oxygen species （ROS） levels in HepG2 cells were observed using a fluorescence inverted microscope. Malondialdehyde （MDA）， glutathione （GSH）， and free Fe2+ content in HepG2 cells were detected using respective kits. The mitochondrial ultrastructure in HepG2 cells was observed by transmission electron microscopy. The expression of ferroptosis-related proteins p53， solute carrier family 7 member 11 （SLC7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， and transferrin receptor 1 （TFR1） in HepG2 cells was detected using Western blot. ResultCompared with the control group， the PPⅡ treatment groups showed significantly decreased survival rate of HepG2 cells in a dose-dependent manner （P<0.01）， significantly reduced number of cell colonies （P<0.01）， significantly shortened scratch healing distance， inverse correlation of the migration distance with drug concentration （P<0.01）， significantly increased LDH leakage in cells （P<0.01）， significantly enhanced relative fluorescence intensity of intracellular ROS， and significantly increased accumulation of lipid peroxide MDA （P<0.01）， decreased intracellular GSH content with increasing drug concentration （P<0.01）， and significantly enhanced fluorescence intensity of FeRhoNox-1 in cells （P<0.01）. Moreover， cells exhibited vacuolation， and mitochondria showed significant shrinkage with reduced or even disappeared cristae. Compared with the results in the control group， the expression of p53， ACSL4， and TFR1 proteins significantly increased， while the expression of SLC7A11 and GPX4 proteins significantly decreased in the PPⅡ treatment groups （P<0.05）. ConclusionIn summary， PPⅡ induces ferroptosis in HepG2 cells by regulating the p53/SLC7A11/GPX4 signaling axis， promoting ACSL4 expression and Fe3+ uptake， leading to an imbalance in the antioxidant system.
