Variability in glucose transporter-1 levels and hexokinase activity in human melanoma.
Melanoma exhibits heterogeneous growth patterns and widely varying sensitivities to multiple treatment modalities. This variability may reflect intrinsic genetic differences in factors giving rise to altered metabolism. Glucose is the primary energy source of tumours, including melanoma, and glucose transporter isoform 1 (Glut-1) and hexokinase are key rate-limiting factors in glucose metabolism. The levels of Glut-1 and total hexokinase activity were measured in 31 melanoma biopsies to determine the extent of tumour-to-tumour variability in these parameters. Relative Glut-1 levels were determined by Western immunoblot analysis using human anti-Glut-1 rabbit polyclonal antibody, and hexokinase activity was measured in the same samples by an enzymatic assay monitoring the reduction in the oxidized form of nicotinamide adenine dinucleotide phosphate (NADP+) (in nmol NADP+ reduced/min per mg protein). All melanomas were from patients who had received no therapy prior to surgery. Immediately after excision, tumour biopsies were disaggregated to single cells by collagenase and DNase and frozen in liquid nitrogen. Thirty human melanomas exhibited a 22-fold variation in levels of Glut-1 and 29 exhibited a nine-fold variation in total cellular hexokinase activity. Glut-1 levels and hexokinase activity were not correlated with one another. The broad range in Glut-1 levels and hexokinase activity observed between melanomas suggests that these glycolytic rate-limiting parameters that influence the rate of glucose metabolism may contribute to the variability in melanoma response to treatment modalities.
Wachsberger
P R
PR
Department of Radiation Oncology, Kimmel Cancer Center, Jefferson Medical College, 111 South 11th Street, Thomas Jefferson University, PA 19107, USA. phyllis.wachsberger@mail.tju.edu
Gressen
E L
EL
Bhala
A
A
Bobyock
S B
SB
Storck
C
C
Coss
R A
RA
Berd
D
D
Leeper
D B
DB
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001706:Biopsy; D015153:Blotting, Western; D003720:Densitometry; D005260:Female; D051272:Glucose Transporter Type 1; D006593:Hexokinase / Q000096:biosynthesis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D008207:Lymphatic Metastasis; D008297:Male; D008545:Melanoma / Q000201:enzymology* / Q000378:metabolism*; D008875:Middle Aged; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis*; D009249:NADP / Q000378:metabolism; D012878:Skin Neoplasms / Q000201:enzymology* / Q000378:metabolism*
D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; C495677:SLC2A1 protein, human; D009249:NADP; D006593:Hexokinase