Effect of Decitabine on DKK1 Gene Demethylation in Leukemia Cells.
- Author:
Feng-Zhi LIU
1
;
Ling HE
2
;
Ji-Shi WANG
2
;
Song ZHANG
3
;
Hong-Qian ZHU
4
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Azacitidine; analogs & derivatives; pharmacology; DNA Methylation; Gene Expression Regulation, Bacterial; Genes, myc; HL-60 Cells; drug effects; Humans; Intercellular Signaling Peptides and Proteins; metabolism; Leukemia, Myeloid, Acute; pathology; RNA, Messenger; Wnt Signaling Pathway; beta Catenin; metabolism
- From: Journal of Experimental Hematology 2016;24(1):56-60
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of decitabine on Dickkopf-1 (DKK1) gene expression level and its downstream Wnt signaling pathway in acute myeloid leukemia (AML) cell line HL-60.
METHODSFlow cytometry and DNA ladder analysis were performed to detect apoptosis in HL-60 cell treated with different concentration of decitabine. Methylation-specific polymerase chain reaction (MS-PCR) was used to examine the methylation status of DKK1 gene. The expressions of mRNA and protein were determined by qRT-PCR and Western blot, respectively.
RESULTSFlow cytometric detection showed that after treating HL-60 cell line with decitabine of different concentrations for 48 h, the early apoptosis of HL-60 cells increased significantly as compared with control group (P < 0.05). DNA ladder analysis showed that the DNA ladder and demethylation of DKK1 gene appeared. RT-PCR and Western blot showed that the expressions of mRNA and protein increased. The protein expressions of β-catenin and C-MYC decreased.
CONCLUSIONThe decitabine can promote the apoptosis of HL-60 cells throngh demethylation of DDK1 gene and inhibition of Wnt signalling pathway.
