title: Des-γ-carboxyl prothrombin induces matrix metalloproteinase activity in hepatocellular carcinoma cells by involving the ERK1/2 MAPK signalling pathway.
abstract: Des-γ-carboxy prothrombin (DCP), an aberrant prothrombin produced by hepatocellular carcinoma (HCC) cells, has been shown to be associated with the biological malignant potential of HCC. The aim of this study was to evaluate the effect of DCP on HCC cell growth and metastasis, and to explore the underlying molecular mechanisms. DCP significantly stimulated HCC cell growth, as measured by cell counting kit-8 assay. Transwell chamber assay showed that DCP increased HCC cell migration through reconstituted extracellular matrix (Matrigel). Gelatin zymography assay and Western blot analysis demonstrated that DCP increased the secretion and expression of matrix metalloproteinase (MMP)-2 and MMP-9 in the supernatant of cultured HCC cells and on tumour cell membranes. DCP was found to bind to the cell surface receptor Met, resulting in Met phosphorylation and subsequent activation of the epidermal growth factor receptor (EGFR). Western blot analysis demonstrated that DCP stimulated a sequential kinase phosphorylation cascade including ERK1/2, MEK1/2 and c-Raf, indicating activation of the extracellular signal-regulated kinase/mitogen activated protein kinase (ERK1/2 MAPK) signalling pathway. Furthermore, blocking ERK1/2 MAPK activation with ERK1/2 inhibitor PD98059 essentially abolished the DCP-induced MMP-2 and MMP-9 activity, confirming the signalling pathway of DCP stimulation. Taken together, these results suggested that DCP stimulates HCC growth and promotes HCC metastasis by increasing the activity of MMP-2 and MMP-9 through activation of the ERK1/2 MAPK signalling pathway.
authors.affiliation: Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan, China.
authors.forename: Pan
authors.initials: P
authors.lastname: Yue
authors.forename: Zu-Hua
authors.initials: ZH
authors.lastname: Gao
authors.forename: Xia
authors.initials: X
authors.lastname: Xue
authors.forename: Shu-Xiang
authors.initials: SX
authors.lastname: Cui
authors.forename: Cui-Rong
authors.initials: CR
authors.lastname: Zhao
authors.forename: Yi
authors.initials: Y
authors.lastname: Yuan
authors.forename: Zhe
authors.initials: Z
authors.lastname: Yin
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Inagaki
authors.forename: Norihiro
authors.initials: N
authors.lastname: Kokudo
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
authors.forename: Xian-Jun
authors.initials: XJ
authors.lastname: Qu
mesh_terms: D015415:Biomarkers; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D066246:ErbB Receptors / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007797:Laminin / Q000737:chemistry; D008113:Liver Neoplasms / Q000201:enzymology* / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009362:Neoplasm Metastasis; D010766:Phosphorylation; D011498:Protein Precursors / Q000494:pharmacology*; D011509:Proteoglycans / Q000737:chemistry; D011516:Prothrombin / Q000494:pharmacology*; D015398:Signal Transduction
chemical_list: D015415:Biomarkers; D004338:Drug Combinations; D007797:Laminin; D011498:Protein Precursors; D011509:Proteoglycans; C059485:matrigel; C008208:acarboxyprothrombin; D011516:Prothrombin; D003094:Collagen; D066246:ErbB Receptors; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9