Home LiteratureArticle Details
PMID: 14762654 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Glucagon-like peptide-1 regulates proliferation and apoptosis via activation of protein kinase B in pancreatic INS-1 beta cells.

Diabetologia ·Vol. 47 ·No. 3 ·2004-03-00 ·Pages 478-487

Wang Q, Li L, Xu E, Wong V, Rhodes C, Brubaker PL

Abstract

The incretin hormone glucagon-like peptide-1 augments islet cell mass in vivo by increasing proliferation and decreasing apoptosis of the beta cells. However, the signalling pathways that mediate these effects are mostly unknown. Using a clonal rat pancreatic beta cell line (INS-1), we examined the role of protein kinase B in mediating beta-cell growth and survival stimulated by glucagon-like peptide-1. Immunoblot analysis was used to detect active (phospho-) and total protein kinase B. Proliferation was assessed using (3)H-thymidine incorporation, while apoptosis was quantitated using 4'-6-diamidino-2-phenylindole staining and APO percentage apoptosis assay. Kinase-dead and wild-type protein kinase B was introduced into cells using adenoviral vectors. Glucagon-like peptide-1 rapidly activated protein kinase B in INS-1 cells (by 2.7+/-0.7-fold, p<0.05). This effect was completely abrogated by inhibition, with wortmannin, of the upstream activator of protein kinase B, phosphatidylinositol-3-kinase. Glucagon-like peptide-1 also stimulated INS-1 cell proliferation in a dose-dependent manner (by 1.8+/-0.5-fold at 10(-7) mol/l, p<0.01), and inhibited staurosporine-induced apoptosis (by 69+/-12%, p<0.05). Both of these effects were also prevented by wortmannin treatment. Ablation of protein kinase B by adenovirus-mediated overexpression of the kinase-dead form of protein kinase Balpha prevented protein kinase B phosphorylation and completely abrogated both cellular proliferation ( p<0.05) and protection from drug-induced cellular death ( p<0.01) induced by glucagon-like peptide-1. These results identify protein kinase B as an essential mediator linking the glucagon-like peptide-1 signal to the intracellular machinery that modulates beta-cell growth and survival.

MeSH Terms
Animals Apoptosis/drug effects Cell Division/drug effects Cell Line Glucagon/pharmacology Glucagon-Like Peptide 1 Insulin/analysis,metabolism Insulin Secretion Islets of Langerhans/cytology,drug effects,physiology Peptide Fragments/pharmacology Protein Precursors/pharmacology Protein Serine-Threonine Kinases/metabolism Proto-Oncogene Proteins/metabolism Proto-Oncogene Proteins c-akt Reverse Transcriptase Polymerase Chain Reaction Staurosporine/pharmacology
Chemicals
Insulin Peptide Fragments Protein Precursors Proto-Oncogene Proteins Glucagon-Like Peptide 1 Glucagon Protein Serine-Threonine Kinases Proto-Oncogene Proteins c-akt Staurosporine
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wang Q
Department of Physiology, University of Toronto, Toronto, Ontario, Canada. | Department of Medicine, Division of Endocrinology and Metabolism, University of Toronto, Toronto, Ontario, Canada. | St Michael's Hospital, University of Toronto, Toronto, Ontario, Canada.
Li L
Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
Xu E
Department of Physiology, University of Toronto, Toronto, Ontario, Canada. | St Michael's Hospital, University of Toronto, Toronto, Ontario, Canada.
Wong V
Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
Rhodes C
Pacific Northwest Research Institute, Seattle, Washington, USA.
Brubaker P L
Department of Physiology, University of Toronto, Toronto, Ontario, Canada. [email protected]. | Department of Medicine, Division of Endocrinology and Metabolism, University of Toronto, Toronto, Ontario, Canada. [email protected]. | Room 3366, Medical Sciences Building, University of Toronto, Toronto, Ontario, M5S 1A8, Canada. [email protected].
References (51)
51 references, click to expand
  1. Endoproteolysis by isolated membrane peptidases reveal metabolic stability of glucagon-like peptide-1 analogs, exendins-3 and -4.
    Exp Clin Endocrinol Diabetes. 2002 May;110(3):113-8 PMID: 12012270
  2. Insulinotropic glucagon-like peptide 1 agonists stimulate expression of homeodomain protein IDX-1 and increase islet size in mouse pancreas.
    Diabetes. 2000 May;49(5):741-8 PMID: 10905482
  3. Pharmacokinetic, insulinotropic, and glucagonostatic properties of GLP-1 [7-36 amide] after subcutaneous injection in healthy volunteers. Dose-response-relationships.
    Diabetologia. 1995 Jun;38(6):720-5 PMID: 7672496
  4. Sustained expression of exendin-4 does not perturb glucose homeostasis, beta-cell mass, or food intake in metallothionein-preproexendin transgenic mice.
    J Biol Chem. 2000 Nov 3;275(44):34471-7 PMID: 10952981
  5. Establishment of 2-mercaptoethanol-dependent differentiated insulin-secreting cell lines.
    Endocrinology. 1992 Jan;130(1):167-78 PMID: 1370150
  6. Regulation of pancreatic beta-cell growth and survival by the serine/threonine protein kinase Akt1/PKBalpha.
    Nat Med. 2001 Oct;7(10):1133-7 PMID: 11590437
  7. Differential effects of subcutaneous GLP-1 on gastric emptying, antroduodenal motility, and pancreatic function in men.
    Proc Assoc Am Physicians. 1997 Jan;109(1):84-97 PMID: 9010920
  8. cAMP-regulated guanine nucleotide exchange factor II (Epac2) mediates Ca2+-induced Ca2+ release in INS-1 pancreatic beta-cells.
    J Physiol. 2001 Oct 15;536(Pt 2):375-85 PMID: 11600673
  9. Mechanisms of mitogenic and anti-apoptotic signaling by glucose-dependent insulinotropic polypeptide in beta(INS-1)-cells.
    J Endocrinol. 2002 Aug;174(2):233-46 PMID: 12176662
  10. Effects of subcutaneous glucagon-like peptide 1 (GLP-1 [7-36 amide]) in patients with NIDDM.
    Diabetologia. 1996 Dec;39(12):1546-53 PMID: 8960841
  11. Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta).
    Science. 2001 Jun 1;292(5522):1728-31 PMID: 11387480
  12. Glucagon-like peptide 1 increases insulin sensitivity in depancreatized dogs.
    Diabetes. 1999 May;48(5):1045-53 PMID: 10331409
  13. Suppression of glucose production by GLP-1 independent of islet hormones: a novel extrapancreatic effect.
    Am J Physiol Endocrinol Metab. 2003 Oct;285(4):E701-7 PMID: 12773303
  14. Glucagon-like peptide-1 promotes DNA synthesis, activates phosphatidylinositol 3-kinase and increases transcription factor pancreatic and duodenal homeobox gene 1 (PDX-1) DNA binding activity in beta (INS-1)-cells.
    Diabetologia. 1999 Jul;42(7):856-64 PMID: 10440129
  15. Glucose-dependent insulinotropic polypeptide activates the Raf-Mek1/2-ERK1/2 module via a cyclic AMP/cAMP-dependent protein kinase/Rap1-mediated pathway.
    J Biol Chem. 2002 Oct 4;277(40):37088-97 PMID: 12138104
  16. Differential signaling of cyclic AMP: opposing effects of exchange protein directly activated by cyclic AMP and cAMP-dependent protein kinase on protein kinase B activation.
    J Biol Chem. 2002 Mar 29;277(13):11497-504 PMID: 11801596
  17. Effect of 6-week course of glucagon-like peptide 1 on glycaemic control, insulin sensitivity, and beta-cell function in type 2 diabetes: a parallel-group study.
    Lancet. 2002 Mar 9;359(9309):824-30 PMID: 11897280
  18. Structure-function of the glucagon receptor family of G protein-coupled receptors: the glucagon, GIP, GLP-1, and GLP-2 receptors.
    Receptors Channels. 2002;8(3-4):179-88 PMID: 12529935
  19. Function of PI3Kgamma in thymocyte development, T cell activation, and neutrophil migration.
    Science. 2000 Feb 11;287(5455):1040-6 PMID: 10669416
  20. Comet assay and DNA flow cytometry analysis of staurosporine-induced apoptosis.
    Cytometry. 1999 Jun 1;36(2):117-22 PMID: 10554159
  21. The proglucagon-derived peptide, glucagon-like peptide-2, is a neurotransmitter involved in the regulation of food intake.
    Nat Med. 2000 Jul;6(7):802-7 PMID: 10888930
  22. Pancreatic beta-cells are rendered glucose-competent by the insulinotropic hormone glucagon-like peptide-1(7-37).
    Nature. 1993 Jan 28;361(6410):362-5 PMID: 8381211
  23. Integrative mitogenic role of protein kinase B/Akt in beta-cells.
    Ann N Y Acad Sci. 2000;921:242-50 PMID: 11193829
  24. Glucagon-like peptide-2 receptor activation engages bad and glycogen synthase kinase-3 in a protein kinase A-dependent manner and prevents apoptosis following inhibition of phosphatidylinositol 3-kinase.
    J Biol Chem. 2002 Jul 12;277(28):24896-906 PMID: 11978789
  25. Protein kinase B is expressed in pancreatic beta cells and activated upon stimulation with insulin-like growth factor I.
    Biochem Biophys Res Commun. 1998 Sep 8;250(1):181-6 PMID: 9735353
  26. Protein kinase B (PKB/Akt)--a key regulator of glucose transport?
    FEBS Lett. 2001 Mar 16;492(3):199-203 PMID: 11257494
  27. Glucagonostatic and insulinotropic action of glucagonlike peptide I-(7-36)-amide.
    Diabetes. 1989 Jul;38(7):902-5 PMID: 2661287
  28. Continuous subcutaneous infusion of glucagon-like peptide 1 lowers plasma glucose and reduces appetite in type 2 diabetic patients.
    Diabetes Care. 1999 Jul;22(7):1137-43 PMID: 10388979
  29. Glucagon-like peptide-1 treatment delays the onset of diabetes in 8 week-old db/db mice.
    Diabetologia. 2002 Sep;45(9):1263-73 PMID: 12242459
  30. Exendin-4 is a high potency agonist and truncated exendin-(9-39)-amide an antagonist at the glucagon-like peptide 1-(7-36)-amide receptor of insulin-secreting beta-cells.
    J Biol Chem. 1993 Sep 15;268(26):19650-5 PMID: 8396143
  31. Glucagon-like peptide 1 receptor signaling influences topography of islet cells in mice.
    Virchows Arch. 2001 Apr;438(4):382-7 PMID: 11355173
  32. Glucose-dependent insulinotropic polypeptide is a growth factor for beta (INS-1) cells by pleiotropic signaling.
    Mol Endocrinol. 2001 Sep;15(9):1559-70 PMID: 11518806
  33. Phosphatidylinositol 3-kinase signaling to Akt mediates survival in isolated canine islets of Langerhans.
    Biochem Biophys Res Commun. 2000 Oct 22;277(2):455-61 PMID: 11032744
  34. Initiation of increased pancreatic islet growth in young normoglycemic mice (Umeå +/?).
    Endocrinology. 1999 Feb;140(2):778-83 PMID: 9927305
  35. Role of endogenous glucagon-like peptide-1 in islet regeneration after partial pancreatectomy.
    Diabetes. 2003 Feb;52(2):365-71 PMID: 12540609
  36. Gastric emptying, glucose responses, and insulin secretion after a liquid test meal: effects of exogenous glucagon-like peptide-1 (GLP-1)-(7-36) amide in type 2 (noninsulin-dependent) diabetic patients.
    J Clin Endocrinol Metab. 1996 Jan;81(1):327-32 PMID: 8550773
  37. PKB/AKT: functional insights from genetic models.
    Nat Rev Mol Cell Biol. 2001 Oct;2(10 ):760-8 PMID: 11584303
  38. Isolation of INS-1-derived cell lines with robust ATP-sensitive K+ channel-dependent and -independent glucose-stimulated insulin secretion.
    Diabetes. 2000 Mar;49(3):424-30 PMID: 10868964
  39. Differential activation of protein kinase B and p70(S6)K by glucose and insulin-like growth factor 1 in pancreatic beta-cells (INS-1).
    J Biol Chem. 2001 Jun 15;276(24):21110-20 PMID: 11274216
  40. Biological actions and therapeutic potential of the glucagon-like peptides.
    Gastroenterology. 2002 Feb;122(2):531-44 PMID: 11832466
  41. Parathyroid hormone and parathyroid hormone-related protein exert both pro- and anti-apoptotic effects in mesenchymal cells.
    J Biol Chem. 2002 May 31;277(22):19374-81 PMID: 11897779
  42. Protein kinase Czeta activation mediates glucagon-like peptide-1-induced pancreatic beta-cell proliferation.
    Diabetes. 2001 Oct;50(10):2237-43 PMID: 11574404
  43. Exendin-4 stimulates both beta-cell replication and neogenesis, resulting in increased beta-cell mass and improved glucose tolerance in diabetic rats.
    Diabetes. 1999 Dec;48(12):2270-6 PMID: 10580413
  44. Biological activities of glucagon-like peptide-1 analogues in vitro and in vivo.
    Biochemistry. 2001 Mar 6;40(9):2860-9 PMID: 11258897
  45. Severe diabetes, age-dependent loss of adipose tissue, and mild growth deficiency in mice lacking Akt2/PKB beta.
    J Clin Invest. 2003 Jul;112(2):197-208 PMID: 12843127
  46. Glucagon-like peptide 1 (GLP-1) as a new therapeutic approach for type 2-diabetes.
    Exp Clin Endocrinol Diabetes. 1997;105(4):187-95 PMID: 9285204
  47. Islet beta cell expression of constitutively active Akt1/PKB alpha induces striking hypertrophy, hyperplasia, and hyperinsulinemia.
    J Clin Invest. 2001 Dec;108(11):1631-8 PMID: 11733558
  48. Glucagon-like peptide-1 receptor signaling modulates beta cell apoptosis.
    J Biol Chem. 2003 Jan 3;278(1):471-8 PMID: 12409292
  49. Protein kinase B/Akt prevents fatty acid-induced apoptosis in pancreatic beta-cells (INS-1).
    J Biol Chem. 2002 Dec 20;277(51):49676-84 PMID: 12393870
  50. Glucagon-like peptide 1 induces pancreatic beta-cell proliferation via transactivation of the epidermal growth factor receptor.
    Diabetes. 2003 Jan;52(1):124-32 PMID: 12502502
  51. The regulation and activities of the multifunctional serine/threonine kinase Akt/PKB.
    Exp Cell Res. 1999 Nov 25;253(1):210-29 PMID: 10579924
Article Info
Journal
Diabetologia
Abbr.
Diabetologia
ISSN
0012-186X
DOI
10.1007/s00125-004-1327-5
Email
[email protected]
Published
2004-03-00
Epub
2004-00-05
Pages
478-487
Language
English
Region
Germany
NLM ID
0006777
Subset
IM
External Links
PubMed source DOI: 10.1007/s00125-004-1327-5 View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2