Glucose regulates both glucose transport and the glucose transporter gene expression in a hamster-derived pancreatic beta-cell line (HIT).
We studied the effect of chronic exposure to high glucose on the glucose transport regulation in hamster pancreatic Beta cells in permanent culture (HIT). Cells were exposed to either 5.5 mmol/l or 16.7 mmol/l glucose for 48 h and then glucose transport was studied by measuring the (3H)-2-deoxyglucose uptake for 5 and 10 min at 37 degrees C. The 2-deoxyglucose uptake was lower in cells pre-exposed to glucose 16.7 mmol/l for 48 h compared to cells pre-exposed to 5.5 (12.0 +/- 1.6 vs 19.1 +/- 1.2 nmol/0.1 mg after 5 min, and 22.2 +/- 2.6 vs 39.0 +/- 2.9 after 10 min respectively, mean +/- SEM, n = 5, p less than 0.01). In order to investigate the mechanism(s) for glucose impairment of glucose transport, we studied the glucose carrier gene expression in the same cells by Northern and slot-blot analysis. When total RNA was extracted from HIT cells cultured at either 5.5 or 16.7 mmol/l glucose and then hybridized to 32P-labelled cDNA probes for the glucose transporter 1, the glucose transporter 2 and beta-actin, a significant reduction of both glucose transporter 1 (-63.9 +/- 4.1%, mean +/- SEM, n = 3) and glucose transporter 2 (-48.9 +/- 3.2%) mRNA was observed in HIT cells cultured with high glucose. In the same experiments no change of beta-actin mRNA was observed, suggesting that the effect of high glucose was specific on the glucose-transporter mRNAs.(ABSTRACT TRUNCATED AT 250 WORDS)
Purrello
F
F
Cattedra di Endocrinologia, University of Catania Medical School, Italy.
Buscema
M
M
Vetri
M
M
Vinci
C
C
Gatta
C
C
Forte
F
F
Rabuazzo
A M
AM
Vigneri
R
R
D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D015152:Blotting, Northern; D002460:Cell Line; D006224:Cricetinae; D015342:DNA Probes; D003847:Deoxyglucose / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D009004:Monosaccharide Transport Proteins / Q000235:genetics*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification
D015342:DNA Probes; D007328:Insulin; D009004:Monosaccharide Transport Proteins; D012313:RNA; D003847:Deoxyglucose; D005947:Glucose