Zebularine induces apoptosis of esophageal cancer cells via demethylation SFRP2/Dkk3 to regulate Wnt/β-catenin signaling pathway
10.16438/j.0513-4870.2020-1828
- VernacularTitle:Zebularine通过SFRP2/Dkk3去甲基化调控Wnt/β-catenin信号通路诱导食管癌细胞凋亡
- Author:
Feng-xia ZHANG
1
;
Jian-dong ZHANG
2
;
Bao-en SHAN
3
;
Jin-xiu CHU
4
Author Information
1. Hebei Key Laboratory for Chronic Diseases of Basic Medical College, North China University of Science and Technology, Tangshan 063200, China
2. Department of Laboratory Medicine, the Third Hospital of Hebei Medical University, Shijiazhuang 050000, China
3. Research Center, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050000, China
4. Hebei Key Laboratory for Chronic Diseases of Basic Medical College, North China University of Science and Technology, Tangshan 063200, China; North China University of Science and Technology, Tangshan 063200, China
- Publication Type:Research Article
- Keywords:
esophageal cancer;
zebularine;
apoptosis;
SFRP2;
Dkk3;
Wnt/β-catenin signaling pathway
- From:
Acta Pharmaceutica Sinica
2021;56(5):1384-1390
- CountryChina
- Language:Chinese
-
Abstract:
To explore the effect and mechanisms of demethylation drug zebularine on esophageal cancer cells apoptosis, ECA109 cells and KYSE170 cells were treated with zebularine at different concentrations (25, 50, 100, 200, and 400 μmol·L-1). The cell viability was measured by CCK-8. Flow cytometry was used to detect the cell apoptosis rate, Western blot was performed to determine the expression of apoptosis protein (Bcl-2, Bax, cleaved-caspase-3, and cleaved-PARP) and Wnt signal pathway molecules (β-catenin, cyclin D1, and c-Myc), real-time quantitative PCR was used to detect the expression level of negative regulatory genes of Wnt signaling pathway, methylation specific PCR (MSP) was used to detect the methylation status of secreted frizzled related protein 2 (SFRP2) and dickkopf 3 (Dkk3) genes. After knockdown of SFRP2 and Dkk3, the effect of zebularine on apoptosis was detected. The studies showed that zebularine could inhibit the activity of ECA109 and KYSE170 cells in a dose-dependent and time-dependent manner; zebularine could induce cell apoptosis, down-regulate the expression of Bcl-2 protein, up-regulate the expression of Bax, cleaved-caspase-3, and cleaved-PARP protein, and inhibit the expression of β-catenin, cyclin D1, and c-Myc protein (P < 0.05); the mRNA expression levels of Dkk3 and SFRP2 were significantly up-regulated by zebularine, while the methylation levels of SFRP2 and Dkk3 promoters were decreased; knockdown of SFRP2 and Dkk3 could reduce the apoptosis induced by zebularine. In summary, zebularine could reduce the methylation level of SFRP2 and Dkk3 gene promoter, promote the expression of SFRP2 and Dkk3 gene, and then induce the apoptosis of esophageal cancer cells by inhibiting Wnt/β-catenin signaling pathway.
