title: Anti-Angiogenic Properties of Cafestol and Kahweol Palmitate Diterpene Esters.
abstract: Epidemiological studies support the association of coffee-specific diterpenes, with various beneficial health effects. Although anti-antiangiogenic properties of free cafestol and kahweol have been recently described, available data regarding their esterified form, in particular palmitate esters as the main diterpene esters present in coffee, are still rare. Given that angiogenesis plays an important role in many pathological conditions, including cancer growth and metastasis, this study aimed to assess and compare the potential anti-angiogenic effects of cafestol palmitate (CP) and kahweol palmitate (KP) in an in vitro angiogenesis model. According to our findings, both compounds inhibited angiogenesis steps on human microvascular endothelial cells (HMVECs), although a more significant effect was observed for KP. Compared to control, HMVECs viability decreased in a dose-dependent manner upon incubation either with CP or KP. Concentrations of 75 and 100 μM of each compound were cytotoxic. Cell proliferation was also dramatically reduced by both diterpene esters at 50 μM, although KP had a stronger inhibitory effect. However, CP and KP did not induce apoptosis on HMVECs. Both compounds reduced cell migration, but this effect was only statistically significant after KP incubation. Inhibition of VEGFR2 expression and its downstream effector Akt, but not Erk, was also observed in CP- and KP-treated HMVECs. These findings were confirmed using ELISA assay for phosphorylated (active) VEGFR-2. Taken together, these data indicate that both CP and KP can be considered potent compounds against angiogenesis-dependent disorders. Our findings further indicate that KP exerts more potent anti-angiogenic effects than CP, in most of assays. J. Cell. Biochem. 117: 2748-2756, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: LEPABE, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
authors.forename: Marzieh
authors.initials: M
authors.lastname: Moeenfard
authors.affiliation: Faculty of Medicine, Department of Biochemistry, University of Porto, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.|I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal.
authors.forename: Alice
authors.initials: A
authors.lastname: Cortez
authors.affiliation: Faculty of Medicine, Department of Biochemistry, University of Porto, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.|I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal.
authors.forename: Vera
authors.initials: V
authors.lastname: Machado
authors.affiliation: Faculty of Medicine, Department of Biochemistry, University of Porto, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.|I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal.
authors.forename: Raquel
authors.initials: R
authors.lastname: Costa
authors.affiliation: Faculty of Medicine, Department of Biochemistry, University of Porto, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.|I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal.
authors.forename: Carla
authors.initials: C
authors.lastname: Luís
authors.affiliation: Faculty of Medicine, Department of Biochemistry, University of Porto, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal.|I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal.|Ciências Químicas e Biomoléculas, Escola Superior de Tecnologias da Saúde do Porto, Instituto Politécnico do Porto, Porto, Portugal.
authors.forename: Pedro
authors.initials: P
authors.lastname: Coelho
authors.affiliation: Faculty of Medicine, Department of Biochemistry, University of Porto, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. raqsoa@med.up.pt.|I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal. raqsoa@med.up.pt.
authors.forename: Raquel
authors.initials: R
authors.lastname: Soares
authors.affiliation: LEPABE, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
authors.forename: Arminda
authors.initials: A
authors.lastname: Alves
authors.affiliation: Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
authors.forename: Nuno
authors.initials: N
authors.lastname: Borges
authors.affiliation: I3S-Instituto de Investigação e Inovação em Saúde, Porto, Portugal.|Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Santos
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004224:Diterpenes / Q000494:pharmacology*; D004952:Esters / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D004224:Diterpenes; D004952:Esters; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C033997:kahweol palmitate; C033998:cafestol palmitate; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: ANGIOGENESIS; COFFEE FAT-SOLUBLE COMPOUNDS; ESTERIFIED DITERPENES; HMVEC