Genetic engineering of glucose-stimulated insulin secretion in Chinese hamster ovary cells.
To engineer an a non-islet cell capable of glucose-stimulated insulin secretion, a chinese hamster ovary cell line (CHO) was transfected with a mammalian expression vector carrying the human insulin cDNA (pCB/hINS). More proinsulin than insulin was released daily by the stably transformed cell line (CHO-INS). Examination of acid-ethanol extracts confirmed that both insulin and proinsulin were stored. Immunohistochemical analysis of the cells also showed that (pro)insulin was stored. Unlike beta cells, CHO-INS cells did not secrete insulin in response to glucose. To investigate this lack of effect, we examined whether transfection of GLUT2 cDNA, which is ordinarily not expressed in CHO-INS cells, would confer glucose-stimulated insulin secretion. Consequently, we have demonstrated that glucose regulated insulin release occurs in the CHO-INS-GLUT2 cell line and that glucose potentiates the insulin secretory response to non-glucose secretagogues.
Kintsurashvili
E
E
Department of Physiology, Faculty of Medicine, University of Toronto, Ontario, Canada.
Zhou
D
D
Wheeler
M B
MB
Vacek
I
I
Sun
A M
AM
D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary; D005818:Genetic Engineering; D005941:Glucokinase / Q000502:physiology; D005947:Glucose / Q000494:pharmacology*; D051273:Glucose Transporter Type 2; D006801:Humans; D007328:Insulin / Q000235:genetics* / Q000378:metabolism*; D000078790:Insulin Secretion; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection
D018076:DNA, Complementary; D051273:Glucose Transporter Type 2; D007328:Insulin; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; D005941:Glucokinase; D005947:Glucose