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GLUT-2 gene transfer into insulinoma cells confers both low and high affinity glucose-stimulated insulin release. Relationship to glucokinase activity.

Publication ,  Journal Article
Ferber, S; BeltrandelRio, H; Johnson, JH; Noel, RJ; Cassidy, LE; Clark, S; Becker, TC; Hughes, SD; Newgard, CB
Published in: J Biol Chem
April 15, 1994

The rat insulinoma cell line RIN 1046-38 loses glucose-stimulated insulin secretion as a function of time in culture. We found that the loss of glucose sensing in these cells was correlated with the loss of expression of GLUT-2 and glucokinase. Stable transfection of RIN cells with a plasmid containing the GLUT-2 cDNA conferred glucose-stimulated insulin release in intermediate but not high passage cells, with the near-maximal 3-fold increase occurring at 50 microM glucose. GLUT-2 expressing cells also exhibited a larger response to the combination of 5 mM glucose + 1 microM forskolin than untransfected cells (7.9 versus 1.6-2.7-fold, respectively). GLUT-2 expressing intermediate passage, but not high passage, RIN cells exhibited a 4-fold increase in glucokinase enzymatic activity relative to nonexpressing controls. Glucokinase activity was also increased by transfer of the GLUT-2 gene into intermediate passage RIN cells via recombinant adenovirus. Preincubation of GLUT-2 expressing intermediate passage RIN cells with 2-deoxyglucose to inhibit low Km hexokinases resulted in a glucose-stimulated insulin secretion response that was shifted toward the physiologic range. These studies indicate that GLUT-2 expression confers both a high and low affinity glucose-stimulated insulin secretion response to intermediate passage RIN cells.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

April 15, 1994

Volume

269

Issue

15

Start / End Page

11523 / 11529

Location

United States

Related Subject Headings

  • Tumor Cells, Cultured
  • Transfection
  • Rats
  • RNA, Messenger
  • Pancreatic Neoplasms
  • Monosaccharide Transport Proteins
  • Methylglucosides
  • Kinetics
  • Insulinoma
  • Insulin Secretion
 

Citation

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Ferber, S., BeltrandelRio, H., Johnson, J. H., Noel, R. J., Cassidy, L. E., Clark, S., … Newgard, C. B. (1994). GLUT-2 gene transfer into insulinoma cells confers both low and high affinity glucose-stimulated insulin release. Relationship to glucokinase activity. J Biol Chem, 269(15), 11523–11529.
Ferber, S., H. BeltrandelRio, J. H. Johnson, R. J. Noel, L. E. Cassidy, S. Clark, T. C. Becker, S. D. Hughes, and C. B. Newgard. “GLUT-2 gene transfer into insulinoma cells confers both low and high affinity glucose-stimulated insulin release. Relationship to glucokinase activity.J Biol Chem 269, no. 15 (April 15, 1994): 11523–29.
Ferber S, BeltrandelRio H, Johnson JH, Noel RJ, Cassidy LE, Clark S, et al. GLUT-2 gene transfer into insulinoma cells confers both low and high affinity glucose-stimulated insulin release. Relationship to glucokinase activity. J Biol Chem. 1994 Apr 15;269(15):11523–9.
Ferber S, BeltrandelRio H, Johnson JH, Noel RJ, Cassidy LE, Clark S, Becker TC, Hughes SD, Newgard CB. GLUT-2 gene transfer into insulinoma cells confers both low and high affinity glucose-stimulated insulin release. Relationship to glucokinase activity. J Biol Chem. 1994 Apr 15;269(15):11523–11529.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

April 15, 1994

Volume

269

Issue

15

Start / End Page

11523 / 11529

Location

United States

Related Subject Headings

  • Tumor Cells, Cultured
  • Transfection
  • Rats
  • RNA, Messenger
  • Pancreatic Neoplasms
  • Monosaccharide Transport Proteins
  • Methylglucosides
  • Kinetics
  • Insulinoma
  • Insulin Secretion