title: Investigation of the molecular response to folate metabolism inhibition.
abstract: We investigated the molecular response to folate metabolism inhibition by exposing human lymphoblast cell lines to the methionine adenosyltransferase inhibitor cycloleucine. We carried out microarray analysis on replicate control and exposed cells by examining 47,000 transcripts on the Affymetrix HG U133 plus 2.0 arrays. We identified 13 genes that we considered reliable responders to cycloleucine treatment: chemokine receptor 3 (CXCR3), prostaglandin-endoperoxide synthase 2, growth arrest-specific 7, reduced folate carrier, klotho beta, early growth response 1, diaphanous homolog 3, prostaglandin D2 synthase (PGDS), butyrophilin-like 9, low-density lipoprotein receptor-related protein 11, chromosome 21 orf15, G-protein-coupled receptor 98 (GPR98) and cystathionine-beta-synthase (CBS). We further demonstrated that four of these genes, CXCR3, PGDS, GPR98 and CBS, consistently responded to cycloleucine treatment in additional experiments over a range of concentrations. We carried out gene-specific DNA methylation analysis on five genes, including CBS, and found no evidence that DNA methylation changes were mediating the gene expression changes observed. Pathway analysis of the microarray data identified four pathways of relevance for response to cycloleucine; the immune response NF-AT signaling pathway was the most statistically significant. Comparison with other gene expression studies focusing on folate deficiency revealed that gene products related to immune cells or the immune response is a common theme. This indicates that apart from their role in the immune response, it is likely that these gene products may also have a role to play in the cellular response to folate status.
authors.affiliation: Nutritional Genomics Group, School of Biotechnology, Dublin City University Dublin 9, Ireland.
authors.forename: Nicola
authors.initials: N
authors.lastname: Carroll
authors.forename: Linda
authors.initials: L
authors.lastname: Hughes
authors.forename: Gráinne
authors.initials: G
authors.lastname: McEntee
authors.forename: Anne
authors.initials: A
authors.lastname: Parle-McDermott
mesh_terms: D000243:Adenosine Deaminase / Q000235:genetics / Q000378:metabolism; D002460:Cell Line / Q000187:drug effects; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics / Q000378:metabolism; D003515:Cycloleucine / Q000494:pharmacology; D003541:Cystathionine beta-Synthase / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004305:Dose-Response Relationship, Drug; D005492:Folic Acid / Q000235:genetics* / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000235:genetics / Q000378:metabolism; D054834:Lipocalins / Q000235:genetics / Q000378:metabolism; D046228:Microarray Analysis; D016601:RNA-Binding Proteins; D054367:Receptors, CXCR3 / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
chemical_list: C417525:ADGRV1 protein, human; C515187:CXCR3 protein, human; D050676:Core Binding Factor Alpha 2 Subunit; C576732:FOXP4 protein, human; D051858:Forkhead Transcription Factors; D054834:Lipocalins; D016601:RNA-Binding Proteins; C493728:RUNX1 protein, human; D054367:Receptors, CXCR3; D043562:Receptors, G-Protein-Coupled; D003515:Cycloleucine; D005492:Folic Acid; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase; D003541:Cystathionine beta-Synthase; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase