Effect of Licochalcone A on Apoptosis in Human Breast Cancer MDA-MB-231 Cells
10.13422/j.cnki.syfjx.20212022
- VernacularTitle:甘草查尔酮A对人乳腺癌MDA-MB-231细胞凋亡的影响
- Author:
Yong-gui YUAN
1
;
Xia-yan ZHANG
1
;
Xiao-jun ZHU
1
;
Wei WANG
2
;
Hai-rong ZENG
2
;
Jia-min LE
1
Author Information
1. Changhai Hospital， Naval Medical University， Shanghai 200433， China
2. Shanghai Putuo District Central Hospital，Shanghai 200062，China
- Publication Type:Research Article
- Keywords:
licochalcone A （LCA）;
breast cancer;
reactive oxygen species;
mitochondrial membrane potential;
endoplasmic reticulum stress
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(20):95-100
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of licochalcone A （LCA） on apoptosis in human breast cancer MDA-MB-231 cells， and to explore its possible mechanism. Method:MDA-MB-231 cells were treated with LCA of different concentrations， and cell counting kit-8 （CCK-8） assay was used to detect the cell viability. The cells were treated with LCA （10， 20， and 40 μmol·L-1） for 24 h， and apoptosis was detected by Annexin V staining with fluorescein isothiocyanate （FITC） and propidium iodide （PI） （Annexin V-FITC/PI）. The level of intracellular reactive oxygen species （ROS） was detected by 2′，7′-dichlorodihydrofluorescein diacetate （DCFA-DA） fluorescent probe. Mitochondrial membrane potential （MMP） was detected by 5， 5′， 6， 6′-tetrachloro-1， 1′， 3， 3′-tetraethyl-imidacarbocyanine （JC-1） fluorescence probe. Western blot was used to detect the expression of cell apoptosis-related proteins， such as B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax）， and endoplasmic reticulum （ER） stress-related proteins， such as C/EBP homologous protein （CHOP）， activating transcription factor 4 （ATF4）， protein kinase R-like ER kinase （PERK）， p-PERK， eukaryotic translation initiation factor 2 alpha （eIF2α）， and p-eIF2α. Result:With the increase in the drug concentration （starting from 5 μmol·L-1）， the cell viability decreased （P<0.05） with IC50 of 19.05 μmol·L-1 as compared with the normal group. Additionally， the apoptosis rates of the LCA groups （10， 20， 40 μmol·L-1） significantly increased （P<0.05）， which reached 30.2% （P<0.05） at LCA concentration of 40 μmol·L-1. LCA （10， 20， and 40 μmol·L-1） decreased the expression of Bcl-2 （P<0.05） and increased Bax expression （P<0.05） in a dose-dependent manner. Besides， the intracellular ROS level was elevated （P<0.05） and mitochondrial MMP was reduced （P<0.05） after LCA （10， 20， and 40 μmol·L-1） treatment in a dose-dependent manner， leading to mitochondrial dysfunction. LCA （10， 20， and 40 μmol·L-1） induced ER stress to up-regulate the expression of CHOP， ATF4， p-PERK， and p-eIF2α （P<0.05） in a dose-dependent manner. Conclusion:LCA can induce MDA-MB-231 cell apoptosis by increasing intracellular ROS level and reducing MMP to trigger mitochondrial dysfunction and ER stress.
