title: Genomic DNA hypomethylation is associated with neural tube defects induced by methotrexate inhibition of folate metabolism.
abstract: DNA methylation is thought to be involved in the etiology of neural tube defects (NTDs). However, the exact mechanism between DNA methylation and NTDs remains unclear. Herein, we investigated the change of methylation in mouse model of NTDs associated with folate dysmetabolism by use of ultraperformance liquid chromatography tandem mass spectrometry (UPLC/MS/MS), liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS), microarray, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and Real time quantitative PCR. Results showed that NTD neural tube tissues had lower concentrations of 5-methyltetrahydrofolate (5-MeTHF, P = 0.005), 5-formyltetrahydrofolate (5-FoTHF, P = 0.040), S-adenosylmethionine (SAM, P = 0.004) and higher concentrations of folic acid (P = 0.041), homocysteine (Hcy, P = 0.006) and S-adenosylhomocysteine (SAH, P = 0.045) compared to control. Methylation levels of genomic DNA decreased significantly in the embryonic neural tube tissue of NTD samples. 132 differentially methylated regions (35 low methylated regions and 97 high methylated regions) were selected by microarray. Two genes (Siah1b, Prkx) in Wnt signal pathway demonstrated lower methylated regions (peak) and higher expression in NTDs (P<0.05; P<0.05). Results suggest that DNA hypomethylation was one of the possible epigenetic variations correlated with the occurrence of NTDs induced by folate dysmetabolism and that Siah1b, Prkx in Wnt pathway may be candidate genes for NTDs.
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, China.
authors.forename: Xiuwei
authors.initials: X
authors.lastname: Wang
authors.affiliation: Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Guan
authors.affiliation: Department of Respiratory, the Second Hospital of Shanxi Medical University, Taiyuan, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, China.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, China.
authors.forename: Yuhu
authors.initials: Y
authors.lastname: Niu
authors.affiliation: Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Wang
authors.affiliation: Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, China; Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Niu
mesh_terms: D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics; D019175:DNA Methylation / Q000187:drug effects* / Q000235:genetics; D004195:Disease Models, Animal; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D005260:Female; D005492:Folic Acid / Q000378:metabolism*; D005493:Folic Acid Antagonists / Q000633:toxicity*; D005494:Folic Acid Deficiency / Q000150:complications; D008727:Methotrexate / Q000633:toxicity*; D051379:Mice; D009436:Neural Tube Defects / Q000209:etiology* / Q000235:genetics / Q000378:metabolism*; D011247:Pregnancy; D017346:Protein Serine-Threonine Kinases; D053719:Tandem Mass Spectrometry; D044767:Ubiquitin-Protein Ligases / Q000235:genetics; D060449:Wnt Signaling Pathway / Q000187:drug effects / Q000235:genetics
chemical_list: D005493:Folic Acid Antagonists; D005492:Folic Acid; C484443:Siah1b protein, mouse; D044767:Ubiquitin-Protein Ligases; C543855:PRKX protein, mouse; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D008727:Methotrexate
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