Barchiesi F Lucchinetti E Zaugg M Ogunshola OO Wright M Meyer M Rosselli M Schaufelberger S Gillespie DG Jackson EK Dubey RK 2010 2-Methoxyestradiol (2-ME; estradiol metabolite) inhibits vascular smooth muscle cell (VSMC) growth and protects against atherosclerosis and vascular injury; however, the mechanisms by which 2-ME induces these actions remain obscure. To assess the impact of 2-ME on biochemical pathways regulating VSMC biology, we used high-density oligonucleotide microarrays to identify differentially expressed genes in cultured human female aortic VSMCs treated with 2-ME acutely (4 hours) or long term (30 hours). Both single gene analysis and Gene Set Enrichment Analysis revealed 2-ME-induced downregulation of genes involved in mitotic spindle assembly and function in VSMCs. Also, Gene Set Enrichment Analysis identified effects of 2-ME on genes regulating cell-cycle progression, cell migration/adhesion, vasorelaxation, inflammation, and cholesterol metabolism. Transcriptional changes were associated with changes in protein expression, including inhibition of cyclin D1, cyclin B1, cyclin-dependent kinase 6, cyclin-dependent kinase 4, tubulin polymerization, cholesterol and steroid synthesis, and upregulation of cyclooxygenase 2 and matrix metalloproteinase 1. Microarray data suggested that 2-ME may activate peroxisome proliferator-activated receptors (PPARs) in VSMCs, and 2-ME has structural similarities with rosiglitazone (PPARγ agonist). However, our finding of weak activation and lack of binding of 2-ME to PPARs suggests that 2-ME may modulate PPAR-associated genes via indirect mechanisms, potentially involving cyclooxygenase 2. Indeed, the antimitogenic effects of 2-ME at concentrations that do not inhibit tubulin polymerization were blocked by the PPAR antagonist GW9662 and the cyclooxygenase 2 inhibitor NS398. Finally, we demonstrated that 2-ME inhibited hypoxia-inducible factor 1α. Identification of candidate genes that are positively or negatively regulated by 2-ME provides important leads to investigate and better understand the mechanisms by which 2-ME induces its vasoprotective actions. application/pdf https://sonar.ch/global/documents/156164 eng info:eu-repo/semantics/altIdentifier/doi/10.1161/HYPERTENSIONAHA.110.152298 info:eu-repo/semantics/altIdentifier/pmid/20921434 info:eu-repo/semantics/openAccess Hypertension (Dallas, Tex. : 1979). - 2010 2-Methoxyestradiol Analysis of Variance Animals Aorta Cells, Cultured Cholesterol Estradiol Female Humans Male Muscle, Smooth, Vascular Myocytes, Smooth Muscle Oligonucleotide Array Sequence Analysis Progesterone Rats Rats, Inbred WKY Testosterone Time Factors Candidate genes and mechanisms for 2-methoxyestradiol-mediated vasoprotection. http://purl.org/coar/resource_type/c_6501