Effect of transforming growth factor-beta1, insulin-like growth factor-I, and hepatocyte growth factor on proteoglycan production and regulation in canine melanoma cell lines.
OBJECTIVE
To identify extracellular proteoglycans produced by canine melanoma cell lines and analyze the effect of transforming growth factor-beta1 (TGF-beta1), insulin-like growth factor-I (IGF-I), and hepatocyte growth factor (HGF) on these proteoglycans.


METHODS
3 canine melanoma cell lines (ie, CML-1, CML-6M, and CML-10c2).


METHODS
Extracellular proteoglycans were analyzed by use of metabolic labeling and western immunoblot analysis. The effect of TGF-beta1 on cell proliferation was determined by incorporation of 5-bromo-2'-deoxyuridine.


RESULTS
The CML-1 and CML-6M melanoma cell lines produced 2 main extracellular proteoglycans. One of them was identified as versican, a proteoglycan found in undifferentiated human melanoma cell lines. The CML-10c2 cells produced a small amount of extracellular proteoglycans. Addition of TGF-beta1 (1.25 to 6.25 ng/ml) increased the release of sulfated proteoglycans into the medium. The TGF-beta1 had mainly a posttranslational effect, because it increased the molecular mass of the sulfated bands. Addition of IGF-I (50 ng/ml) slightly increased production of proteoglycans in the CML-6M cell line, whereas HGF (50 ng/ml) did not have any effect on proteoglycan production.


CONCLUSIONS
The proteoglycan content and response toTGF-beta1 treatment for CML-1 and CML-6M canine melanoma cell lines are similar to that for undifferentiated human melanoma cell lines. In contrast, CML-10c2 cells produced a low amount of proteoglycans with high molecular weight. Because these extracellular proteoglycans are involved in the control of cell adhesion, proliferation, and migration, they may play an important role in the progression of melanomas in dogs.
Serra
Montserrat
M
Departament de Bioquímica i Biologia Molecular, Facultat de Veterinária, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Pastor
Josep
J
Domenzain
Clelia
C
Bassols
Anna
A
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002455:Cell Division / Q000502:physiology; D011508:Chondroitin Sulfate Proteoglycans / Q000096:biosynthesis / Q000378:metabolism; D004283:Dog Diseases / Q000378:metabolism*; D004285:Dogs; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D037181:Lectins, C-Type; D008545:Melanoma / Q000378:metabolism* / Q000662:veterinary*; D011509:Proteoglycans / Q000096:biosynthesis* / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1; D014407:Tumor Cells, Cultured; D053675:Versicans
D011508:Chondroitin Sulfate Proteoglycans; D037181:Lectins, C-Type; D011509:Proteoglycans; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; C508322:VCAN protein, human; D053675:Versicans; D017228:Hepatocyte Growth Factor; D007334:Insulin-Like Growth Factor I