Effects of arsenic and its main metabolites on A549 cell apoptosis and the expression of pro-apoptotic genes Bad and Bik.
10.3760/cma.j.cn121094-20201208-00676
- Author:
Qian ZHOU
1
;
Jin Yao YIN
1
;
Jing Wen TAN
1
;
Shu Ting LI
1
;
Cheng Lan JIANG
1
;
Yue Feng HE
1
Author Information
1. School of Public Health, Kunming Medical University, Kunming 650500, China.
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Arsenic;
Bad;
Bik;
Dimethyl arsonic acid (DMA);
Metabolomics;
Monomethyl arsonic acid (MMA);
Sodium arsenite
- MeSH:
A549 Cells;
Adenosine Diphosphate Ribose/pharmacology*;
Apoptosis;
Apoptosis Regulatory Proteins;
Arsenic;
Arsenites;
Cacodylic Acid/pharmacology*;
Caspase 3;
Caspases/pharmacology*;
Cytochromes c/pharmacology*;
Humans;
Mitochondrial Proteins/pharmacology*;
Poisons;
Propidium/pharmacology*;
RNA, Messenger;
Sincalide/pharmacology*;
Sodium Compounds;
bcl-Associated Death Protein/metabolism*
- From:
Chinese Journal of Industrial Hygiene and Occupational Diseases
2022;40(9):661-667
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of arsenic and its main metabolites on the apoptosis of human lung adenocarcinoma cell line A549 and the expression of pro-apoptotic genes Bad and Bik. Methods: In October 2020, A549 cells were recovered and cultured, and the cell viability was detected by the cell counting reagent CCK-8 to determine the concentration and time of sodium arsenite exposure to A549. The study was divided into NaAsO(2) exposure groups and metobol: le expoure groups: the metabolite comparison groups were subdivided into the control group, the monomethylarsinic acid exposure group (60 μmol/L) , and the dimethylarsinic acid exposure group (60 μmol/L) ; sodium arsenite dose groups were subdivided into 4 groups: control group (0) , 20, 40, 60 μmol/L sodium arsenite NaAsO(2). Hoechst 33342/propidium iodide double staining (Ho/PI) was used to observe cell apoptosis and real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of Bad and Bik mRNA in cells after exposure. Western blotting was used to detect the protein expressions of Bad, P-Bad-S112, Bik, cleaved Bik and downstream proteins poly ADP-ribose polymerase PARP1 and cytochrome C (Cyt-C) , using spectrophotometry to detect the activity changes of caspase 3, 6, 8, 9. Results: Compared with the control group, the proportion of apoptotic cells in the 20, 40, and 60 μmol/L NaAsO(2) dose groups increased significantly (P<0.01) , and the expression levels of Bad, Bik mRNA, the protein expression levels of Bad, P-Bad-S112, Bik, cleaved Bik, PARP1, Cyt-C were increased (all P<0.05) , and the activities of Caspase 3, 6, 8, and 9 were significantly increased with significantly differences (P<0.05) . Compared with the control group, the expression level of Bad mRNA in the DMA exposure group (1.439±0.173) was increased with a significant difference (P=0.024) , but there was no significant difference in the expression level of Bik mRNA (P=0.788) . There was no significant differences in the expression levels of Bad and Bik mRNA in the poison groups (P=0.085, 0.063) . Compared with the control group, the gray values of proteins Bad, Bik, PARP1 and Cyt-C exposed to MMA were 0.696±0.023, 0.707±0.014, 0.907±0.031, 1.032±0.016, and there was no significant difference between the two groups (P=0.469, 0.669, 0.859, 0.771) ; the gray values of proteins Bad, Bik, PARP1 and Cyt-C exposed to DMA were 0.698±0.030, 0.705±0.022, 0.908±0.015, 1.029±0.010, and there was no difference between the two groups (P=0.479, 0.636, 0.803, 0.984) . Conclusion: Sodium arsenite induces the overexpression of Bad and Bik proteins, initiates the negative feedback regulation of phosphorylated Bad and the degradation of Bik, activates the downstream proteins PARP1, Cyt-C and Caspase pathways, and mediates the apoptosis of A549 cells.
