Summary
Glucose-stimulated insulin secretion, glucose transport, glucose phosphorylation and glucose utilization have been characterized in the insulinoma cell line MIN6, which is derived from a transgenic mouse expressing the large T-antigen of SV40 in pancreatic beta cells. Glucose-stimulated insulin secretion occurred progressively from 5 mmol/l glucose, reached the maximal level approximately seven-fold above the basal level at 25 mmol/l, and remained at this level up to 50 mmol/l. Glucose transport was very rapid with the half-maximal uptake of 3-O-methyl-d-glucose being reached within 15 s at 22 °C. Glucose phosphorylating activity in the cell homogenate was due mainly to glucokinase; the Vmax value of glucokinase activity was estimated to be 255±37 nmol·h−1·mg protein−1, constituting approximately 80% of total phosphorylating activity, whereas hexokinase activity constituted less than 20%. MIN6 cells exhibited mainly the high Km component of glucose utilization with a Vmax of 289±18 nmol·h−1·mg protein−1. Thus, glucose utilization quantitatively and qualitatively reflected glucose phosphorylation in MIN6 cells. In contrast, MIN7 cells, which exhibited only a small increase in insulin secretion in response to glucose, had 4.7-fold greater hexokinase activity than MIN6 cells with a comparable activity of glucokinase. These characteristics in MIN6 cells are very similar to those of isolated islets, indicating that this cell line is an appropriate model for studying the mechanism of glucose-stimulated insulin secretion in pancreatic beta cells.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Miyazaki J-I, Araki K, Yamato E et al. (1990) Establishment of a pancreatic beta cell line that retains glucose-inducible insulin secretion: special reference to expression of glucose transporter isoforms. Endocrinology 127: 126–132
Halban PA, Praz GA, Wollheim CB (1983) Abnormal glucose metabolism accompanies failure of glucose to stimulate insulin release from a rat pancreatic cell line (RINm5 F) Biochem J 212: 439–443
Giroix M-H, Sener A, Dufrane SP, Malaisse-Lagae F, Malaisse WJ (1985) Glucose metabolism in insulin-producing tumoral cells. Arch Biochem Biophys 241: 561–570
Clark SA, Burnham BL, Chick WL (1990) Modulation of glucose-induced insulin secretion from a rat clonal β-cell line. Endocrinology 127: 2779–2788
Hamaguchi K, Gaskins HR, Leiter EH (1991) NIT-1, a pancreatic β-cell line established from a transgenic NOD/Lt mouse. Diabetes 40: 842–849
Efrat S, Linde S, Kofod H et al. (1988) Beta-cell lines derived from transgenic mice expressing a hybrid insulin gene-oncogene. Proc Natl Acad Sci USA 85: 9037–9041
Whitesell RR, Powers AC, Regan DM, Abumrad NA (1991) Transport and metabolism of glucose in an insulin-secreting cell line, βTC-1. Biochemistry 30: 11560–11566
D'Ambra R, Manju S, Efrat S, Starr RG, Fleischer N (1990) Regulation of insulin secretion from β-cell lines derived from transgenic mice insulinomas resembles that of normal β-cells. Endocrinology 126: 2815–2822
Gilligan A, Jewett L, Simon D et al. (1989) Functional pancreatic β-cell line from SV40 T-antigen transgenic mouse. Diabetes 38: 1056–1062
Wang SY (1989) The acute effects of glucose on the insulin biosynthetic-secretory pathway in a Simian Virus 40-transformed hamster pancreatic islet β-cell line. Endocrinology 124: 1980–1987
Asfari M, Janjic D, Meda P, Li G, Halban PA, Wollheim CB (1992) Establishment of 2-mercaptoethanol-dependent differentiated insulin-secreting cell lines. Endocrinology 130: 167–178
Meglasson MD, Manning CD, Najafi H, Matschinsky FM (1986) Glucose transport by radiation-induced insulinoma and clonal pancreatic β-cells. Diabetes 35: 1340–1344
Malaisse WJ, Giroix M-H, Malaisse-Lagae F, Sener A (1986) 3-O-methyl-d-glucose transport in tumoral insulin-producing cells. Am J Physiol 251: C841-C846
Trautmann ME, Wollheim CB (1987) Characterization of glucose transport in an insulin-secreting cell line. Biochem J 242: 625–630
Ashcroft SJH, Stubbs M (1987) The glucose sensor in HIT cells is the glucose transporter. FEBS Lett 219: 311–315
Shimizu T, Knowles BB, Matschinsky FM (1988) Control of glucose phosphorylation and glucose usage in clonal insulinoma cells. Diabetes 37: 563–568
Grodsky GM, Batts AA, Bannett LL, Vcella C, McWilliams NB, Smith DF (1963) Effects of carbohydrates on secretion of insulin from isolated rat pancreas. Am J Physiol 205: 638–644
Trus MD, Zawalich WS, Burch PT, Berner DK, Weill VA, Matschinsky FM (1981) Regulation of glucose metabolism in pancreatic islets. Diabetes 30: 911–922
Unger R (1991) Diabetic hyperglycemia: link to impaired glucose transport in pancreatic beta cells. Science (Wash.DC) 251: 1200–1205
Epstein PN, Boschero AC, Atwater I, Xingang C, Overbeek PA (1992) Expression of yeast hexokinase in pancreatic β cells of transgenic mice reduces blood glucose, enhances insulin secretion, and decreases diabetes. Proc Natl Acad Sci USA 89: 12038–12042
Johnson JH, Ogawa A, Chen L et al. (1990) The high Km glucose transporter of islets of Langerhans is functionally similar to the low affinity transporter of liver and has an identical primary sequence. J Biol Chem 265: 6548–6551
Ashcroft SJH, Weerasinghe LCC, Bassett JM, Randle PJ (1972) The pentose cycle and insulin release in mouse pancreatic islets. Biochem J 126: 525–532
Lernmark Å (1974) The preparation of, and studies on, free cell suspensions from mouse pancreatic islets. Diabetologia 10: 431–438
Giroix M-H, Sener A, Pipeleers DG, Malaisse WJ (1984) Hexose metabolism in pancreatic islets: inhibition of hexokinase. Biochem J 223: 447–453
Tiedge M, Lenzen S (1991) Regulation of glucokinase and GLUT-2 glucose transporter gene expression in pancreatic beta cells. Biochem J 279: 899–901
Hughes SD, Quaade C, Milburn JL, Cassidy L, Newgard CB (1991) Expression of normal and novel glucokinase mRNAs in anterior pituitary and islet cells. J Biol Chem 266: 4521–4530
Vionnet N, Stoffel M, Takeda J et al. (1992) Nonsense mutation in the glucokinase gene causes early-onset non-insulin-dependent diabetes mellitus. Nature 356: 721–722
Katagiri H, Asano T, Ishihara H et al. (1992) Nonsense mutation of glucokinase gene in late-onset non-insulin-dependent diabetes mellitus. Lancet 340: 1316–1317
Sener A, Kawazu S, Hutton JC et al. (1978) The stimulus-secretion coupling of glucose-stimulated insulin release: effect of exogenous pyruvate on islet function. Biochem J 176: 217–232
Lindström P (1984) Kinetics of 5-hydroxytryptophan potentiation of glucose-induced insulin release. Acta Endocrinol 106: 248–253
Eizirik DL, Strandell E, Sandler S (1988) Culture of mouse pancreatic islets in different glucose concentrations modifies B cell sensitivity to streptozotocin. Diabetologia 31: 168–174
Pipeleers DG, Schuit FC, In't Veld PA et al. (1985) Interplay of nutrients and hormones in the regulation of insulin release. Endocrinology 117: 824–833
Lernmark Å, Sehlin J, Täljedal I-B (1975) The use of dispersed pancreatic islet cells in measurements of transmembrane transport. Anal Biochem 63: 73–79
Thorens B, Sarkar HK, Kaback HR, Lodish HF (1988) Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney, and β-pancreatic islet cells. Cell 55: 281–290
Inagaki N, Yasuda K, Inoue G et al. (1992) Glucose as regulator of glucose transport activity and glucose-transporter mRNA in hamster beta-cell line. Diabetes 41: 592–597
Yasuda K, Yamada Y, Inagaki N et al. (1992) Expression of GLUT1 and GLUT2 glucose transporter isoforms in rat islets of Langerhans and their regulation by glucose. Diabetes 41: 76–81
Tal M, Thorens B, Surana M et al. (1992) Glucose transporter isotypes switch in T-antigen transformed pancreatic β cells growing in culture and in mice. Mol Cell Biol 12: 422–432
Katagiri H, Asano T, Ishihara H et al. (1992) Replacement of intracellular C-terminal domain of GLUT1 glucose transporter with that of GLUT2 increases Vmax and Km of transport activity. J Biol Chem 267: 22550–22555
Salas J, Salas M, Vinuela E, Sols A (1965) Glucokinase of rabbit liver: purification and properties. J Biol Chem 240: 1014–1018
Purrello F, Buscema M, Rabuazzo AM et al. (1993) Glucose modulates glucose transporter affinity, glucokinase activity, and secretory response in rat pancreatic β-cells. Diabetes 42: 199–205
Davagnino J, Ureta T (1980) The identification of extrahepatic “glucokinase” as N-acetylglucosamine kinase. J Biol Chem 255: 2633–2636
Meglasson MD, Burch PT, Berner DK, Najafi H, Vogin AP, Matschinsky FM (1983) Chromatographic resolution and kinetic characterization of glucokinase from islets of Langerhans. Proc Natl Acad Sci USA 80: 85–89
Gorus FK, Malaisse WJ, Pipeleers DG (1984) Differences in glucose handling by pancreatic A- and B-cells. J Biol Chem 259: 1196–1200
Matschinsky FM, Pagliara AS, Stillings SN, Hover BA (1976) Glucose and ATP levels in pancreatic islet tissue of normal and diabetic rats. J Clin Invest 58: 1193–1200
Reyes A, Cardenas ML (1984) All hexokinase isoenzymes coexist in rat hepatocytes. Biochem J 221: 303–309
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ishihara, H., Asano, T., Tsukuda, K. et al. Pancreatic beta cell line MIN6 exhibits characteristics of glucose metabolism and glucose-stimulated insulin secretion similar to those of normal islets. Diabetologia 36, 1139–1145 (1993). https://doi.org/10.1007/BF00401058
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00401058