Isolation of genes involved in pancreas regeneration by subtractive hybridization
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Jong-Ho Choi
Abstract
The deterioration of β cells in the pancreas is a crucial factor in the progression of diabetes mellitus; therefore, the recovery of β cells is of vital importance for effective diabetic therapeutic strategies. Partially pancreatectomized rats have been used for the investigation of pancreatic regeneration. Because it was determined that tissue extract from the partially-dissected pancreas induces pancreatic differentiation in embryonic stem cells, paracrine factors were thought to be involved in the regeneration. In this study, we screened for genes that had higher mRNA levels 2 days after 60%-pancreatectomy. The genes were isolated using subtractive hybridization and DNA sequencing. Twelve genes (adipsin, Aplp2, Clu, Col1a2, Glul, Krt8, Lgmn, LOC299907, LOC502894, Pla2g1b, Reg3α and Xbp1) were identified, and RT-PCR and real-time PCR analyses were performed to validate their expression levels. Among the genes identified, three genes (Glul, Lgmn and Reg3a) were selected for further analyses. Assays revealed that Glul and Reg3α enhance cell growth. Glul, Lgmn and Reg3α change the expression level of islet marker genes, where NEUROD, NKX2.2, PAX4 and PAX6 are up-regulated and somatostatin is down-regulated. Thus, we believe that Glul, Lgmn and Reg3a can serve as novel targets in diabetes mellitus genetic therapy.
©2010 by Walter de Gruyter Berlin New York
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Artikel in diesem Heft
- Review
- Molecular mechanisms of signal transduction via adiponectin and adiponectin receptors
- Genes and Nucleic Acids
- Isolation of genes involved in pancreas regeneration by subtractive hybridization
- Wild type p53-dependent transcriptional upregulation of cathepsin L expression is mediated by C/EBPα in human glioblastoma cells
- Protein Structure and Function
- Epstein-Barr virus lytic cycle activation alters proteasome subunit expression in Burkitt's lymphoma cells
- Dentipain, a Streptococcus pyogenes IdeS protease homolog, is a novel virulence factor of Treponema denticola
- Membranes, Lipids, Glycobiology
- Resveratrol treatment induces redox stress in red blood cells: a possible role of caspase 3 in metabolism and anion transport
- Cell Biology and Signaling
- Activation of the plasma kallikrein-kinin system on human lung epithelial cells
- Antiproliferative effects induced by guanine-based purines require hypoxanthine-guanine phosphoribosyltransferase activity
- Phosphorylation of caldesmon by myosin light chain kinase increases its binding affinity for phosphorylated myosin filaments
- Proteolysis
- Effects of magnesium ions on recombinant human furin: selective activation of hydrolytic activity upon substrates derived from virus envelope glycoprotein