2018 ACVIM Forum Research Abstract Program: Seattle, Washington, June 14 - 15, 2018

Canine melanoma cells (MCM-N1) were cultured in Dulbecco';s Modified Eagle';s medium containing 1 g/L glucose and 10% fetal bovine serum. Cell growth was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction assay. The cells proliferated for three days in a time-dependent manner. When treated with WZB-117, a pan-glucose transporter inhibitor, the cell growth was clearly attenuated. We observed, by RT-PCR, that mRNAs of glucose transporter subtypes GLUT1 and 3 were dominantly expressed in canine melanoma. The protein expression of GLUT1 and 3 was confirmed by Western blotting. We, then, investigated the contribution of GLUT subtypes on the cell growth by subtype-specific knockdown of GLUT1 and 3. When the cells were transfected with GLUT1 and 3 siRNAs, the cell growth was inhibited. We also observed that the expression of proliferation marker Ki67 was inhibited in GLUT3 knockdown cells, whereas the inhibition was less in GLUT1 knockdown cells, hence suggesting that GLUT3 dominantly contributes to cell growth in canine melanoma.