title: Melatonin Regulates Angiogenic Factors under Hypoxia in Breast Cancer Cell Lines.
abstract: Angiogenesis is the process of new blood vessel formation, regulated by a number of pro- and antiangiogenic factors and usually begins in response to hypoxia. Exogenous administration of melatonin has shown numerous anti-tumor effects and appears to inhibit tumor angiogenesis. However, many factors involved in the anti-angiogenic effect of melatonin are still under investigation. Here, we evaluate the effects of melatonin on cell viability and expression of angiogenic factors in MCF-7 and MDA-MB-231 breast cancer cells under hypoxic conditions. Cell viability was investigated by MTT and gene and protein expression of the hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF-A) were verified by qPCR and immunocytochemistry after melatonin treatment (1 mM) under hypoxic conditions. Additionally, a protein array with 20 different cytokines/factors was performed on tumor cell lysates. The results showed that 1 mM of melatonin reduced the viability of MCF-7 and MDA-MB-231 cells (p < .05). This treatment also decreased both gene and protein expression of HIF-1α and VEGF-A under hypoxic conditions (p < .05). Among the proteins evaluated by protein array, melatonin treatment during hypoxia reduced VEGF-C, VEGFR receptors (VEGFR2 and VEGFR3), matrix metalloproteinase 9 (MMP9) and Angiogenin in MCF-7 cells. In MDA-MB-231 cells, a significant decrease was observed in VEGFR2, epidermal growth factor receptor (EGFR) and Angiogenin (p < .05). Taken together, these results showed that melatonin acts in the regulation of angiogenic factors in breast tumor cells and suggests an anti-angiogenic activity, particularly under hypoxic conditions.
authors.forename: Bruna Victorasso
authors.initials: BV
authors.lastname: Jardim-Perassi
authors.forename: Mateus Repolês
authors.initials: MR
authors.lastname: Lourenço
authors.forename: Gabriel Mandarini
authors.initials: GM
authors.lastname: Doho
authors.forename: Ingrid Helen
authors.initials: IH
authors.lastname: Grígolo
authors.forename: Gabriela Bottaro
authors.initials: GB
authors.lastname: Gelaleti
authors.forename: Lívia Carvalho
authors.initials: LC
authors.lastname: Ferreira
authors.forename: Thaiz Ferraz
authors.initials: TF
authors.lastname: Borin
authors.forename: Marina Gobbe
authors.initials: MG
authors.lastname: Moschetta
authors.affiliation: Laboratório de Investigação Molecular no Câncer (LIMC), Faculdade de Medicina de São José do Rio Preto/FAMERP. Avenida Brigadeiro Faria Lima, 5416, Vila São Pedro, 15090-000 São José do Rio Preto, SP, Brazil. debora.zuccari@famerp.br.
authors.forename: Debora Aparecida
authors.initials: DA
authors.lastname: Pires de Campos Zuccari
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000098:blood supply*; D015687:Cell Hypoxia / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D005260:Female; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D061986:MCF-7 Cells; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008550:Melatonin / Q000494:pharmacology*; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D040321:Vascular Endothelial Growth Factor Receptor-3
chemical_list: D020533:Angiogenesis Inhibitors; D000975:Antioxidants; D016207:Cytokines; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C467484:VEGFA protein, human; C467486:VEGFC protein, human; D042461:Vascular Endothelial Growth Factor A; D042582:Vascular Endothelial Growth Factor C; C512478:EGFR protein, human; D066246:ErbB Receptors; C581309:FLT4 protein, human; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2; D040321:Vascular Endothelial Growth Factor Receptor-3; C046366:angiogenin; D012259:Ribonuclease, Pancreatic; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9; D008550:Melatonin