title: Effects of melatonin on HIF-1α and VEGF expression and on the invasive properties of hepatocarcinoma cells.
abstract: Liver cancer is the sixth most commonly occurring cancer globally, and the main histological type is hepatocellular carcinoma. This type of neoplasia has a poor prognosis due to a high rate of recurrence and intrahepatic metastasis, which are closely are closely associated with the angiogenic process. Vascular endothelial growth factor (VEGF), which is under the control of hypoxia inducible factor-1α (HIF-1α), stimulates the proliferation of endothelial cells and increases cell permeability, promoting the growth, spread and metastasis of tumors. Melatonin, the main hormone secreted by the pineal gland, may have a significant role in tumor suppression and has demonstrated antiangiogenic and antimetastatic effects. The aim of the present study was to analyze the cell viability, migration and invasion, as well as the expression of proangiogenic proteins VEGF and HIF-1α, in HepG2 hepatocarcinoma cells, following treatment with melatonin. Cells were cultured and cell viability was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of proangiogenic proteins VEGF and HIF-1α, under conditions of normoxia and hypoxia, was verified using immunocytochemistry and quantified by densitometry. The analysis of the processes of cell migration and invasion was performed in a Boyden chamber. The MTT assay revealed a reduction in cell viability (P=0.018) following treatment with 1 mM melatonin for 24 h. The expression of proangiogenic proteins VEGF and HIF-1α was reduced in cells treated with 1 mM melatonin for 24 h in normoxic (P<0.001) and hypoxic (P<0.001) conditions, compared with the control group and with induced hypoxia alone. The rate of cell migration and invasion was additionally reduced in cells treated with 1 mM melatonin for 48 h when compared with the control group (P=0.496). The results of the present study suggest that melatonin may have an antiproliferative, antiangiogenic and antimetastatic role in hepatocarcinoma cells and may present a novel therapeutic option for the treatment of liver cancer.
authors.affiliation: Laboratory of Molecular Investigation of Cancer (LIMC), Faculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, São Paulo, SP 15090-000, Brazil.
authors.forename: Jucimara
authors.initials: J
authors.lastname: Colombo
authors.affiliation: Department of Molecular Biology, Faculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, São Paulo, SP 15090-000, Brazil.
authors.forename: João Marcos Wolf
authors.initials: JM
authors.lastname: Maciel
authors.affiliation: Laboratory of Molecular Investigation of Cancer (LIMC), Faculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, São Paulo, SP 15090-000, Brazil; Postgraduate Program in Genetics, Universidade Estadual Paulista "Júlio de Mesquita Filho" - UNESP/IBILCE, São José do Rio Preto, São Paulo, SP 15054-000, Brazil.
authors.forename: Lívia Carvalho
authors.initials: LC
authors.lastname: Ferreira
authors.affiliation: Department of Surgery, Faculty of Medicine of São José do Rio Preto, São José do Rio Preto São Paulo, SP 15090-000, Brazil.
authors.forename: Renato Ferreira
authors.initials: RF
authors.lastname: DA Silva
authors.affiliation: Laboratory of Molecular Investigation of Cancer (LIMC), Faculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, São Paulo, SP 15090-000, Brazil; Postgraduate Program in Genetics, Universidade Estadual Paulista "Júlio de Mesquita Filho" - UNESP/IBILCE, São José do Rio Preto, São Paulo, SP 15054-000, Brazil; Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto, São José do Rio Preto São Paulo, SP 15090-000, Brazil.
authors.forename: Debora Aparecida Pires
authors.initials: DA
authors.lastname: Zuccari
keywords: HepG2; hypoxia-inducible factor 1α; invasion assay; liver cancer; melatonin; vascular endothelial growth factor