title: Induction of a trophoblast-like phenotype by hydralazine in the p19 embryonic carcinoma cell line.
abstract: Chemicals that affect cellular differentiation through epigenetic mechanisms have potential utility in treating a wide range of diseases. Hydralazine decreases DNA methylation in some cell types but its effect on differentiation has not been well explored. After five days of exposure to hydralazine, P19 embryocarcinoma cells displayed a giant cell morphology and were binucleate, indicative of a trophoblast-like morphology. Other trophoblast-like properties included the intermediary filament Troma-1/cytokeratin 8 and the transcription factor Tead4. A decrease in CpG methylation at three sites in the TEAD4 promoter and the B1 repeated sequence was observed. Knocking down expression of Tead4 with siRNA blocked the increase in Troma-1/cytokeratin 8 and over expression of Tead4 induced the expression of Troma-1/cytokeratin 8. Cells treated for 5days with hydralazine were no longer capable of undergoing retinoic acid-mediated neuronal differentiation. An irreversible loss of the pluripotent transcription factor Oct-4 was observed following hydralazine exposure. In summary, hydralazine induces P19 cells to assume a trophoblast-like phenotype by upregulating Tead4 expression through a mechanism involving DNA demethylation.
authors.affiliation: Division of Toxicological Sciences, Department of Environmental Health Sciences, Johns Hopkins Bloomberg School of Public Health and Hugo Moser Research Institute at Kennedy Krieger, Baltimore, MD 21205, USA. odriscoll@kennedykrieger.org
authors.forename: Cliona M
authors.initials: CM
authors.lastname: O'Driscoll
authors.forename: Jonathan B
authors.initials: JB
authors.lastname: Coulter
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Bressler
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D054278:Embryonal Carcinoma Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D006830:Hydralazine / Q000494:pharmacology*; D007124:Immunoenzyme Techniques; D053533:Keratin-8 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000091262:TEA Domain Transcription Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D014327:Trophoblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D014407:Tumor Cells, Cultured
chemical_list: D004268:DNA-Binding Proteins; D053533:Keratin-8; D009124:Muscle Proteins; D012333:RNA, Messenger; D000091262:TEA Domain Transcription Factors; C101204:Tead4 protein, mouse; D014157:Transcription Factors; D006830:Hydralazine; D014212:Tretinoin