title: The acquisition of an insulin-secreting phenotype by HGF-treated rat pancreatic ductal cells (ARIP) is associated with the development of susceptibility to cytokine-induced apoptosis.
abstract: The elucidation of mechanisms regulating the regeneration and survival of pancreatic beta cells has fundamental implications in the cell therapy of type 1 diabetes. The present study had the following three aims: 1. to investigate whether pancreatic ductal epithelial cells can be induced to differentiate into insulin-producing cells by exposing them to hepatocyte growth factor (HGF); 2. to characterize some of the molecular events leading to their differentiation toward a beta-cell-like phenotype; 3. to evaluate the susceptibility of newly differentiated insulin-secreting cells to cytokine-induced apoptosis, a mechanism of beta-cell destruction occurring in type 1 diabetes. We demonstrated that HGF-treated rat pancreatic ductal cell line (ARIP) cells acquired the capability to transcribe the insulin gene and translate its counterpart protein. HGF-treated cells also exhibited a glucose-dependent capability to secrete insulin into the cultured medium. Expression analysis of some of the genes regulating pancreatic beta-cell differentiation revealed a time-dependent transcription of neurogenin-3 and Neuro-D in response to HGF. Finally, we determined the susceptibility to proinflammatory cytokine (PTh1)-induced apoptosis by incubating HGF-treated and untreated ARIP cells with a cocktail of interleukin-1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). Such treatment induced apoptotic death, as determined by the TUNEL technique, in about 40% of HGF-treated, insulin-secreting ARIP cells, while untreated ARIP cells were resistant to PTh1-induced apoptosis. In conclusion, we showed that HGF promotes the differentiation of ARIP cells into pancreatic beta-cell-like cells, and that the differentiation toward an insulin-secreting phenotype is associated with the appearance of susceptibility to cytokine-induced apoptosis.
authors.affiliation: Department of Clinical Sciences, University La Sapienza, Rome, Italy. emanuela.anastasi@uniroma1.it
authors.forename: E
authors.initials: E
authors.lastname: Anastasi
authors.forename: C
authors.initials: C
authors.lastname: Santangelo
authors.forename: A
authors.initials: A
authors.lastname: Bulotta
authors.forename: F
authors.initials: F
authors.lastname: Dotta
authors.forename: B
authors.initials: B
authors.lastname: Argenti
authors.forename: C
authors.initials: C
authors.lastname: Mincione
authors.forename: A
authors.initials: A
authors.lastname: Gulino
authors.forename: M
authors.initials: M
authors.lastname: Maroder
authors.forename: R
authors.initials: R
authors.lastname: Perfetti
authors.forename: U
authors.initials: U
authors.lastname: Di Mario
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D051792:Basic Helix-Loop-Helix Proteins; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D000078790:Insulin Secretion; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000502:physiology; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010183:Pancreatic Ducts / Q000166:cytology*; D010641:Phenotype; D051381:Rats; D000071656:Receptor, Notch3; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D051880:Receptors, Notch; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D016207:Cytokines; D005947:Glucose; D017228:Hepatocyte Growth Factor; D007328:Insulin; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D000071656:Receptor, Notch3; D011956:Receptors, Cell Surface; D051880:Receptors, Notch; D014157:Transcription Factors; C498695:NEUROG3 protein, human; C498696:Neurog3 protein, rat; C498556:Notch3 protein, rat; C093564:Neurogenic differentiation factor 1