In vivo overexpression of tumstatin domains by tumor cells inhibits their invasive properties in a mouse melanoma model.
Our previous studies demonstrated that a synthetic peptide encompassing residues 185-203 of the noncollagenous (NC1) domain of the alpha3 chain of type IV collagen, named tumstatin, inhibits in vitro melanoma cell proliferation and migration. In the present study, B16F1 melanoma cells were stably transfected to overexpress the complete tumstatin domain (Tum 1-232) or its C-terminal part, encompassing residues 185-203 (Tum 183-232). Tumstatin domain overexpression inhibited B16F1 in vitro cell proliferation, anchorage-independent growth, and invasive properties. For studying the in vivo effect of overexpression, representative clones were subcutaneously injected into the left side of C57BL6 mice. In vivo tumor growth was decreased by -60% and -56%, respectively, with B16F1 cells overexpressing Tum 1-232 or Tum 183-232 compared to control cells. This inhibitory effect was associated with a decrease of in vivo cyclin D1 expression. We also demonstrated that the overexpression of Tum 1-232 or Tum 183-232 induced an in vivo down-regulation of proteolytic cascades involving matrix metalloproteinases (MMPs), especially the production or activation of MMP-2, MMP-9, MMP-13, as well as MMP-14. The plasminogen activation system was also altered in tumors with a decrease of urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA) and a strong increase of plasminogen activator inhibitor-1 (PAI-1). Collectively, our results demonstrate that tumstatin or its C-terminal antitumor fragment, Tum 183-232, inhibits in vivo melanoma progression by triggering an intracellular transduction pathway, which involves a cyclic AMP (cAMP)-dependent mechanism.
Pasco
Sylvie
S
Laboratory of Medical Biochemistry and Molecular Biology, Faculty of Medicine, CNRS UMR 6198, F-51095 Reims, France.
Ramont
Laurent
L
Venteo
Lydie
L
Pluot
Michel
M
Maquart
François-Xavier
FX
Monboisse
Jean-Claude
JC
D000818:Animals; D001324:Autoantigens / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D024141:Collagen Type IV / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D015316:Genetic Therapy; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009361:Neoplasm Invasiveness / Q000517:prevention & control*; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics; D010960:Plasminogen Activators / Q000235:genetics; D017434:Protein Structure, Tertiary; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection
D001324:Autoantigens; D024141:Collagen Type IV; D017395:Plasminogen Activator Inhibitor 1; C057112:type IV collagen alpha3 chain; D000242:Cyclic AMP; D045726:Metalloproteases; D010960:Plasminogen Activators