Effect of 5-aza-CdR demethylation on expression of SHP-1 and C-kit genes in leukemia HL-60 cells.
10.7534/j.issn.1009-2137.2014.06.014
- Author:
Zhen MENG
1
;
Ying-Hua LI
2
;
Dong-Mei WANG
2
;
Jian-Min LUO
3
Author Information
1. Department of Hemotology, Harrison International Peace Hospital, Hengshui 053000, Hebei Province, China. E-mail: mengzhen1978@163.com.
2. Department of Hemotology, Harrison International Peace Hospital, Hengshui 053000, Hebei Province, China.
3. Department of Hemotology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China.
- Publication Type:Journal Article
- MeSH:
Azacitidine;
pharmacology;
DNA Methylation;
drug effects;
HL-60 Cells;
Humans;
Leukemia;
genetics;
metabolism;
Protein Tyrosine Phosphatase, Non-Receptor Type 6;
genetics;
Proto-Oncogene Proteins c-kit;
genetics;
RNA, Messenger
- From:
Journal of Experimental Hematology
2014;22(6):1572-1576
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the expression level of SHP-1 and C-kit genes in acute leukemia HL-60 cells and effect of 5-aza-CdR demethylation on expression of SHP-1 and C-kit genes. RT-PCR was used to detect the mRNA expression level of SHP-1 and C-kit mRNA in HL-60 cells of the drug-treated group and control group.The methylation specific PCR (MSP) was applied to measure the methylation status of SHP-1 and C-kit genes in HL-60 cells.The results showed that after being treated with 5-aza-CdR, the recovery of SHP-1 gene expression was observed in HL-60 cells in which SHP-1 mRNA originally was not expressed. Meanwhile, the high expression level of C-kit mRNA in HL-60 cells was decreased. When HL-60 cells were treated with 0, 0.5, 1.0, 2.0 µmol/L 5-aza-CdR, the demethylation effect was enhanced, the expression of SHP-1 mRNA displayed an ascending tendency, and the expression of C-kit mRNA showed an descending tendency in dose-dependent manner (P < 0.05) . It is concluded that the absence of SHP-1 mRNA expression in HL-60 cells and recovery of expression after treatment with 5-aza-CdR suggest that the hypermethylation of SHP-1 gene relates with pathogenesis of leukemia, and the abnormal increase of C-kit mRNA expression maybe exist in formation of leukemia. The effect of 5-aza-CdR on expression of SHP-1 and C-kit shows dose-dependency, the higher the 5-aza-CdR concentration, the higher the SHP-1 expression and the lower the C-kit expression, moreover, the effect of 5-aza-CdR shows time-dependency in specific concentration.The SHP-1 mRNA expression negatively correlates with C-kit mRNA expression, suggesting that the decrease or absence of SHP-1 expression in leukemia cells weakens the negative regulation on C-kit signaling pathway, thus plays a role in the formation of leukemia.
