Document Detail


Cultured pancreatic ductal cells undergo cell cycle re-distribution and beta-cell-like differentiation in response to glucagon-like peptide-1.
MedLine Citation:
PMID:  12459036     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The intestinal hormone glucagon-like peptide-1 (GLP-1) has been shown to promote an increase in pancreatic beta-cell mass via proliferation of islet cells and differentiation of non-insulin-secreting cells. In this study, we have characterized some of the events that lead to the differentiation of pancreatic ductal cells in response to treatment with human GLP-1. Rat pancreatic ductal (ARIP) cells were cultured in the presence of GLP-1 and analyzed for cell counting, cell cycle distribution, expression of cyclin-dependent-kinase (Cdk) inhibitors, transcription of beta-cell-specific genes, loss of ductal-like phenotype and acquisition of beta-cell-like gene expression profile. Exposure of ARIP cells to 10 nM GLP-1 induced a significant reduction in the cell replication rate and a significant decrease in the percentage of cells in S phase of the cell cycle. This was associated with an increase in the number of cells in G0-G1 phase and a reduction of cells in G2-M phase. Western blot analysis for the Cdk inhibitors, kinase inhibitor protein 1 (p27(Kip1)) and Cdk-interacting protein 1 (p21(Cip1)), demonstrated a significant increase in p27(Kip1) and p21(Cip1) levels within the first 24 h from the beginning of GLP-1 treatment. As cells slowed down their proliferation rate, GLP-1 also induced a time-dependent expression of various beta-cell-specific mRNAs. The glucose transporter GLUT-2 was the first of those factors to be expressed (24 h treatment), followed by insulin (44 h) and finally by the enzyme glucokinase (56 h). In addition, immunocytochemistry analysis showed that GLP-1 induced a time-dependent down-regulation of the ductal marker cytokeratin-20 (CK-20) and a time-dependent induction of insulin expression. Finally, GLP-1 promoted a glucose-dependent secretion of insulin, as demonstrated by HPLC and RIA analyses of the cell culture medium. The present study has demonstrated that GLP-1 induces a cell cycle re-distribution with a decrease in cell proliferation rate prior to promoting the differentiation of cells towards an endocrine-like phenotype.
Authors:
A Bulotta; H Hui; E Anastasi; C Bertolotto; L G Boros; U Di Mario; R Perfetti
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of molecular endocrinology     Volume:  29     ISSN:  0952-5041     ISO Abbreviation:  J. Mol. Endocrinol.     Publication Date:  2002 Dec 
Date Detail:
Created Date:  2002-12-02     Completed Date:  2003-10-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8902617     Medline TA:  J Mol Endocrinol     Country:  England    
Other Details:
Languages:  eng     Pagination:  347-60     Citation Subset:  IM    
Affiliation:
Cedars-Sinai Medical Center, Los Angeles, California 90048, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle / drug effects*
Cell Cycle Proteins / metabolism
Cell Differentiation / drug effects*
Cells, Cultured
Cyclin-Dependent Kinase Inhibitor p21
Cyclin-Dependent Kinase Inhibitor p27
Cyclins / metabolism
Flow Cytometry
Gene Expression Regulation / drug effects
Glucagon / pharmacology*
Glucagon-Like Peptide 1
Glucokinase / metabolism
Glucose / pharmacology
Glucose Transporter Type 2
Humans
Immunohistochemistry
Insulin / analysis,  secretion
Monosaccharide Transport Proteins / metabolism
Pancreatic Ducts / cytology*,  drug effects*
Peptide Fragments / pharmacology*
Protein Precursors / pharmacology*
Rats
Tumor Suppressor Proteins / metabolism
Chemical
Reg. No./Substance:
0/CDKN1A protein, human; 0/Cdkn1a protein, rat; 0/Cdkn1b protein, rat; 0/Cell Cycle Proteins; 0/Cyclin-Dependent Kinase Inhibitor p21; 0/Cyclins; 0/Glucose Transporter Type 2; 0/Monosaccharide Transport Proteins; 0/Peptide Fragments; 0/Protein Precursors; 0/Tumor Suppressor Proteins; 11061-68-0/Insulin; 147604-94-2/Cyclin-Dependent Kinase Inhibitor p27; 50-99-7/Glucose; 89750-14-1/Glucagon-Like Peptide 1; 9007-92-5/Glucagon; EC 2.7.1.2/Glucokinase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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