DNA methylation-dependent suppression of HIF1A in an immature hematopoietic cell line HMC-1.
The HMC-1 cell line represents the phenotype of immature mast cells. The HIF1A gene product HIF-1alpha plays key roles in maintaining oxygen homeostasis in eukaryotic organisms and is involved in many processes, including immune response and hematopoiesis. In this study we investigated HIF1A expression in HMC-1 immature hematopoietic cells and CD34+ hematopoietic progenitors. HMC-1 cells exhibited exceptionally low levels of HIF1A expression compared to other cell lines as determined by real-time PCR, and multipotent CD34+ hematopoietic progenitors in bone marrow exhibited significantly lower levels of HIF1A mRNA compared to mature blood cells in peripheral blood. We searched for the mechanisms responsible for suppression of HIF1A expression in HMC-1 cells and obtained evidence for a DNA methylation-dependent process. In vitro methylation of the HIF1A promoter resulted in a decrease in its transcriptional activity and the level of DNA methylation in the HIF1A promoter region in analyzed cell lines was negatively correlated with HIF1A expression. Furthermore, the DNA demethylating agent 5'-azacytidine increased HIF1A expression, and MeCP2 protein was preferentially associated with the HIF1A promoter in vivo. In conclusion, we report that the HIF1A gene in HMC-1 immature hematopoietic cells is suppressed by a process dependent on DNA methylation, and we present evidence indicating downregulation of HIF1A expression in multipotent CD34+ hematopoietic progenitors.
Walczak-Drzewiecka
Aurelia
A
Laboratory of Cellular Immunology, Institute of Medical Biology, Polish Academy of Sciences, Poland.
Ratajewski
Marcin
M
Pułaski
Łukasz
Ł
Dastych
Jarosław
J
D001374:Azacitidine / Q000494:pharmacology; D002460:Cell Line; D045744:Cell Line, Tumor; D019175:DNA Methylation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics
D004791:Enzyme Inhibitors; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D001374:Azacitidine