title: [Effect of 5-aza-CdR demethylation on expression of SHP-1 and C-kit genes in leukemia HL-60 cells].
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.
authors.affiliation: Department of Hemotology, Harrison International Peace Hospital, Hengshui 053000, Hebei Province, China. E-mail: mengzhen1978@163.com.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Meng
authors.affiliation: Department of Hemotology, Harrison International Peace Hospital, Hengshui 053000, Hebei Province, China.
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Li
authors.affiliation: Department of Hemotology, Harrison International Peace Hospital, Hengshui 053000, Hebei Province, China.
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Wang
authors.affiliation: Department of Hemotology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China.
authors.forename: Jian-Min
authors.initials: JM
authors.lastname: Luo
mesh_terms: D001374:Azacitidine / Q000494:pharmacology*; D019175:DNA Methylation / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000235:genetics* / Q000378:metabolism; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000235:genetics*; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics*; D012333:RNA, Messenger
chemical_list: D012333:RNA, Messenger; D019009:Proto-Oncogene Proteins c-kit; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D001374:Azacitidine