title: Potent angiogenic inhibition effects of deacetylated chitohexaose separated from chitooligosaccharides and its mechanism of action in vitro.
abstract: This study was performed to demonstrate the effects of deacetylated chitohexaose (hexamer) separated from a chitooligosaccharide (COS) mixture on tumor angiogenesis and its mechanism of action. Five fractions from dimer to hexamer were separated by a linear gradient solution of HCl on a cation-exchange resin. Then HCl was removed from the fractions by a charcoal column. The purity of the five fractions was analyzed by HPLC and the molecular masses were analyzed by MALDI-TOFMS. The hexamer expressed an inhibitory influence on CAM angiogenesis in a dose-dependent manner at concentrations of 6.25-50microg/egg. On further investigation, we found that the hexamer had no toxic effect on normal ECV304 cells, but could inhibit the proliferation and migration of tumor-induced ECV304 cells in a dose-dependent manner. The mechanism was demonstrated through the detection of mRNA expression of VEGF, MMP-9, TIMP-1, TIMP-2, and uPA by RT-PCR, which showed that the hexamer down-regulated the VEGF and uPA mRNA expressions in ECV304 cells, but up-regulated the TIMP-1 mRNA expression.
authors.affiliation: Dalian Institute of Chemical Physics, Chinese Academy of Science, China.
authors.forename: Chuannan
authors.initials: C
authors.lastname: Xiong
authors.forename: Haige
authors.initials: H
authors.lastname: Wu
authors.forename: Peng
authors.initials: P
authors.lastname: Wei
authors.forename: Ma
authors.initials: M
authors.lastname: Pan
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Tuo
authors.forename: Isao
authors.initials: I
authors.lastname: Kusakabe
authors.forename: Yuguang
authors.initials: Y
authors.lastname: Du
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D015394:Molecular Structure; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009844:Oligosaccharides / Q000737:chemistry* / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
chemical_list: D009844:Oligosaccharides; D019715:Tissue Inhibitor of Metalloproteinase-1; D042461:Vascular Endothelial Growth Factor A; D019716:Tissue Inhibitor of Metalloproteinase-2; C050523:chitohexaose; D048271:Chitosan; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9