title: Effects of Gekko sulfated polysaccharide-protein complex on human hepatoma SMMC-7721 cells: inhibition of proliferation and migration.
abstract: OBJECTIVE
  Gekko swinhonis Guenther has been used as an anti-cancer drug in traditional Chinese medicine for hundreds of years. Here we investigated the structural characterization and anti-cancer effects of sulfated polysaccharide-protein complex (GSPP) isolated from Gekko swinhonis Guenther.
  
  
  METHODS
  The structure of GSPP was characterized by high performance liquid chromatography, gas chromatography, gas chromatography-mass spectrometry, beta-elimination reaction, and NMR spectroscopy. SMMC-7721 cells were used to assess the influence of GSPP on hepatocellular carcinoma. Cell proliferation and survival was determined by trypan blue exclusion assay. Cell migration was performed by wound-healing and transwell assay. The secretion of IL-8 was detected by an enzyme-linked immunosorbent assay kit. Flow cytometry was used to analyze intracellular calcium concentration, as well as cell cycle distribution and apoptosis. Confocal microscopy was used to assess the localization and configuration of actin filaments.
  
  
  RESULTS
  GSPP was chemically characterized as a sulfated polysaccharide-protein complex with O-glycopeptide linkages. Our results showed that GSPP inhibited the proliferation of SMMC-7721 cells and blocked cells in the S phase. No direct toxicity against cells was observed. Furthermore, GSPP inhibited the migration of SMMC-7721 cells with the reduction of intracellular calcium. Actin filaments were polymerized and accumulated in the cytoplasm of the treated cells, whereas the secretion of IL-8 was not significantly changed after GSPP exposure.
  
  
  CONCLUSIONS
  We describe an identified sulfated polysaccharide-protein complex, and demonstrate its direct effect on hepatocellular carcinoma cell migration via calcium-mediated regulation of the actin cytoskeleton reorganization.
authors.affiliation: Hemorheology Center, Department of Medical Physics, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.forename: Wei-Juan
authors.initials: WJ
authors.lastname: Yao
authors.forename: Xiao-Lan
authors.initials: XL
authors.lastname: Zhang
authors.forename: Xiao-Qiang
authors.initials: XQ
authors.lastname: Han
authors.forename: Xiao-Yuan
authors.initials: XY
authors.lastname: Qu
authors.forename: Wei-Bo
authors.initials: WB
authors.lastname: Ka
authors.forename: Da-Gong
authors.initials: DG
authors.lastname: Sun
authors.forename: Xiong-Zhi
authors.initials: XZ
authors.lastname: Wu
authors.forename: Zong-Yao
authors.initials: ZY
authors.lastname: Wen
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use*; D002118:Calcium / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000556:secondary; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D006023:Glycoproteins / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D016209:Interleukin-8 / Q000378:metabolism; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000556:secondary; D008116:Lizards; D008516:Medicine, Chinese Traditional; D009361:Neoplasm Invasiveness; D009945:Organotherapy; D011134:Polysaccharides / Q000494:pharmacology / Q000627:therapeutic use*; D016196:S Phase / Q000187:drug effects
chemical_list: D000199:Actins; D000970:Antineoplastic Agents; D006023:Glycoproteins; D016209:Interleukin-8; D011134:Polysaccharides; C519079:gepsin; D002118:Calcium