Differential gene expression in well-regulated and dysregulated pancreatic beta-cell (MIN6) sublines.
To identify genes involved in regulated insulin secretion, we have established and characterized two sublines derived from the mouse pancreatic beta-cell line MIN6, designated B1 and C3. They have a similar insulin content, but differ in their secretory properties. B1 responded to glucose in a concentration- and cell confluence-dependent manner, whereas C3 did not. B1 cells were stimulated by phorbol 12-myristate 13-acetate, leucine, arginine, glibenclamide, isobutylmethylxanthine, and KCl, whereas C3 did not respond (leucine, arginine, and glibenclamide) or responded to a lesser extent (isobutylmethylxanthine, phorbol 12-myristate 13-acetate, and KCl). Although intracellular Ca(2+) rose in response to glucose in B1 but not C3 cells, KCl increased intracellular Ca(2+) in a similar manner in both sublines. GLUT-1, GLUT-2, Kir6.2, and SUR1 expression was not significantly different between B1 and C3 cells, whereas E-cadherin was more abundantly expressed in B1 cells. A more complete list of differentially expressed genes was established by suppression subtractive hybridization and high density (Affymetrix) oligonucleotide microarrays. Genes were clustered according to known or putative function. Those involved in metabolism, intracellular signaling, cytoarchitecture, and cell adhesion are of potential interest. These two sublines should be useful for identification of the genes and mechanisms involved in regulated insulin secretion of the pancreatic beta-cell.
Lilla
Valérie
V
Louis-Jeantet Research Laboratories, University Medical Center, 1211 Geneva 4, Switzerland. valerie.lilla@medecine.unige.ch
Webb
Gene
G
Rickenbach
Katharina
K
Maturana
Andres
A
Steiner
Donald F
DF
Halban
Philippe A
PA
Irminger
Jean-Claude
JC
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002452:Cell Count; D002460:Cell Line; D005786:Gene Expression Regulation / Q000502:physiology; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D016900:Neurofilament Proteins / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis
D007328:Insulin; D016900:Neurofilament Proteins; C044882:neurofilament protein L; C061195:neurofilament protein M; D005947:Glucose; D002118:Calcium