title: Pemetrexed alters folate phenotype and inflammatory profile in EA.hy 926 cells grown under low-folate conditions.
abstract: Elevated homocysteine is a risk marker for several major human pathologies. Emerging evidence suggests that perturbations of folate/homocysteine metabolism can directly modify production of inflammatory mediators. Pemetrexed acts by inhibiting thymidylate synthetase (TYMS), dihydrofolate reductase (DHFR), and glycinamide ribonucleotide formyltransferase (GARFT). EA.hy 926 cells grown under low ("Lo") and high ("Hi") folate conditions were treated with pemetrexed. The concentrations of several intracellular folate derivatives were measured using LC-MRM/MS. Lo cells had lower total folate concentrations and a different distribution of the intracellular folate derivatives than Hi cells. Treatment with pemetrexed caused a decrease in individual folate analytes. Microarray analysis showed that several genes were significantly up or down-regulated in pemetrexed treated Lo cells. Several of the significantly up-regulated transcripts were inflammatory. Changes in transcript levels of selected targets, including C3, IL-8, and DHFR, were confirmed by quantitative RT-PCR. C3 and IL-8 transcript levels were increased in pemetrexed-treated Lo cells relative to Lo controls; DHFR transcript levels were decreased. In Lo cells, IL-8 and C3 protein concentrations were increased following pemetrexed treatment. Pemetrexed drug treatment was shown in this study to have effects that lead to an increase in pro-inflammatory mediators in Lo cells. No such changes were observed in Hi cells, suggesting that pemetrexed could not modify the inflammatory profile in the context of cellular folate sufficiency.
authors.affiliation: Centers for Cancer Pharmacology, Pharmacogenetics, and Excellence in Environmental Toxicology, Department of Pharmacology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104-6160, USA.
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Hammons
authors.forename: Carolyn M
authors.initials: CM
authors.lastname: Summers
authors.forename: Jeanine
authors.initials: J
authors.lastname: Jochems
authors.forename: Jasbir S
authors.initials: JS
authors.lastname: Arora
authors.forename: Suhong
authors.initials: S
authors.lastname: Zhang
authors.forename: Ian A
authors.initials: IA
authors.lastname: Blair
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Whitehead
mesh_terms: D002460:Cell Line; D018932:Chemokine CCL2 / Q000235:genetics; D003176:Complement C3 / Q000235:genetics; D005492:Folic Acid / Q000378:metabolism*; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D005971:Glutamates / Q000494:pharmacology*; D006147:Guanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D016209:Interleukin-8 / Q000235:genetics; D000068437:Pemetrexed; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics
chemical_list: C111835:CCL2 protein, human; D018932:Chemokine CCL2; D003176:Complement C3; D005493:Folic Acid Antagonists; D005971:Glutamates; D016209:Interleukin-8; D012333:RNA, Messenger; D000068437:Pemetrexed; D006147:Guanine; D005492:Folic Acid; D013762:Tetrahydrofolate Dehydrogenase