title: The Involvement of Hepatocyte Growth Factor-MET-Matrix Metalloproteinase 1 Signaling in Bladder Cancer Invasiveness and Proliferation. Effect of the MET Inhibitor, Cabozantinib (XL184), on Bladder Cancer Cells.
abstract: OBJECTIVE
  To clarify the invasive mechanisms of muscle-invasive bladder cancer (BCa) would be useful for the determination of appropriate treatment strategies. We previously showed that hepatocyte growth factor (HGF)-MET signaling is correlated with invasiveness of BCa cells. Here, we investigated the effects of the MET inhibitor, cabozantinib (XL184), on BCa cells.
  
  
  METHODS
  We first conducted Western blot analysis to investigate MET expression in BCa cell lines. Next, we examined the effect of cabozantinib on their proliferation and invasive abilities using MTT and Matrigel invasion assays, respectively. Invasion assays were performed using the xCELLigence system. Additionally, to investigate the biological function of HGF-MET signaling, we analyzed gene expression profiles and performed real-time polymerase chain reaction analyses of 5637 cells that were cultivated with or without HGF stimulation, with or without cabozantinib.
  
  
  RESULTS
  MET was highly expressed in 4 of 5 BCa cell lines, and 5637 and T24 cells showed especially high protein expression of MET. Cabozantinib suppressed cell proliferation and invasion (cell index; mock, 1.49 vs HGF, 2.26 vs HGF + XL184, 1.47, P < .05). Gene expression profile analysis indicated that matrix metalloproteinase 1 (MMP1) was significantly elevated at the mRNA level with addition of HGF. Moreover, cabozantinib suppressed HGF-induced MMP1 expression in 5637 T24 cells.
  
  
  CONCLUSIONS
  These data indicate that cabozantinib suppressed MMP1 expression by blocking HGF-MET signaling and that HGF-MET-MMP1 signaling is involved in the invasiveness and proliferation of BCa cells. These results suggest that cabozantinib might prove useful for future treatment of muscle-invasive BCa.
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Terumichi
authors.initials: T
authors.lastname: Shintani
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Kusuhara
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Daizumoto
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Tsogt-Ochir
authors.initials: TO
authors.lastname: Dondoo
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yamamoto
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Hidehisa
authors.initials: H
authors.lastname: Mori
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Fukawa
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan. Electronic address: nakatsuji@kameihospital.com.
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Nakatsuji
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Tomoharu
authors.initials: T
authors.lastname: Fukumori
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Department of Urology, Institute of Biomedical Sciences, Tokushima University, Graduate School, Tokushima, Japan.
authors.forename: Hiroomi
authors.initials: H
authors.lastname: Kanayama
mesh_terms: D000813:Anilides / Q000494:pharmacology*; D015153:Blotting, Western; D002452:Cell Count; D045744:Cell Line, Tumor / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis / Q000235:genetics*; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000473:pathology; D040081:Protein Array Analysis; D019859:Proto-Oncogene Proteins c-met / Q000096:biosynthesis / Q000235:genetics*; D011725:Pyridines / Q000494:pharmacology*; D012334:RNA, Neoplasm / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism
chemical_list: D000813:Anilides; C066855:HGF protein, human; D011725:Pyridines; D012334:RNA, Neoplasm; C558660:cabozantinib; D017228:Hepatocyte Growth Factor; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met; D020794:Receptor Protein-Tyrosine Kinases; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1