title: Fucoidan/FGF-2 induces angiogenesis through JNK- and p38-mediated activation of AKT/MMP-2 signalling.
abstract: Angiogenesis is an important biological process in tissue development and repair. Fucoidan has previously been shown to potentiate in vitro tube formation in the presence of basic fibroblast growth factor (FGF-2). However, the underlying molecular mechanism remains largely unknown. This study was designed to investigate the action of fucoidan in angiogenesis in human umbilical vein endothelial cells (HUVECs) and to explore fucoidan-signalling pathways. First, we evaluated the effect of fucoidan on cell proliferation. Matrigel-based tube formation and wound healing assays were performed to investigate angiogenesis. Matrix metalloproteinase-2 (MMP-2) mRNA expression and activity levels were analysed by reverse transcription polymerase chain reaction (RT-PCR) and zymography, respectively. Additionally, phosphorylation of mitogen-activated protein kinases (MAPKs) and protein kinase B (AKT) was detected by Western blot. The results indicate that fucoidan treatment significantly increased cell proliferation in the presence of FGF-2. Moreover, compared to the effect of FGF-2 alone, fucoidan and FGF-2 had a greater effect on tube formation and cell migration, and this effect was found to be synergistic. Furthermore, fucoidan enhanced the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38, and AKT. MMP-2 activation was also significantly increased. Specific inhibitors of p38 (SB203580) and JNK (SP600125) inhibited tube formation and wound healing, while an ERK inhibitor (PD98059) did not. MMP-2 activation and AKT phosphorylation were also attenuated and associated with the suppression of p38 and JNK phosphorylation, but not with that of ERK. These results indicate that fucoidan, in the presence of FGF-2, induces angiogenesis through AKT/MMP-2 signalling by activating p38 and JNK. These findings provide basic molecular information on the effect of fucoidan on angiogenesis in the presence of FGF-2.
authors.affiliation: Wonkwang Bone Regeneration Research Institute, Wonkwang University, Iksan, Jeonbuk 570-749, Republic of Korea; Bonecell Biotech Inc., 77, Dunsan-dong, Seo-gu, Daejeon 302-830, Republic of Korea.
authors.forename: Beom Su
authors.initials: BS
authors.lastname: Kim
authors.affiliation: Bonecell Biotech Inc., 77, Dunsan-dong, Seo-gu, Daejeon 302-830, Republic of Korea.
authors.forename: Ji-Yun
authors.initials: JY
authors.lastname: Park
authors.affiliation: Wonkwang Bone Regeneration Research Institute, Wonkwang University, Iksan, Jeonbuk 570-749, Republic of Korea.
authors.forename: Hyo-Jin
authors.initials: HJ
authors.lastname: Kang
authors.affiliation: Department of Microbiology, School of Medicine, Wonkwang University, Iksan, Jeonbuk 570-749, Republic of Korea.
authors.forename: Hyung-Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Wonkwang Bone Regeneration Research Institute, Wonkwang University, Iksan, Jeonbuk 570-749, Republic of Korea; Bonecell Biotech Inc., 77, Dunsan-dong, Seo-gu, Daejeon 302-830, Republic of Korea. Electronic address: omslee@wku.ac.kr.
authors.forename: Jun
authors.initials: J
authors.lastname: Lee
mesh_terms: D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010766:Phosphorylation; D011134:Polysaccharides / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D011134:Polysaccharides; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; C007789:fucoidan; D051057:Proto-Oncogene Proteins c-akt; D048051:p38 Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D020778:Matrix Metalloproteinase 2
keywords: AKT; Angiogenesis; Fucoidan; MMP-2; Mitogen-activated protein kinase (MAPK)