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Gastron stimulates migration of AGS gastric epithelial cells
GASTRON
STIMULATES
MIGRATION
OF AGS GASTRIC
EPITHELIAL
CELLS
Noble, Peter-John Mantyla; Dockray, Graham J; Varro, Andrea Physiological Laboratory, University of Liverpool, United Kingdom Gastric epithelial cells migrate to form gastric tubules, during epithelial restitution and during tumour invasion. Gastrin regulates the organisation of the gastric epithelium and here we report it stimulates migration of AGS gastric epithelial cells stably transfected with the gastrin-CCKB receptor (AGS-Gr). AGS-Gr cell migration was quantified in a wound healing assay on confluent monolayers and using time lapse videomicroscopy. Cell-cell interactions were investigated by co-culturing AGS-Gr cells with AGS cells lacking the gastrin receptor but constitutively expressing green fluorescent protein (AGS-GFP cells). Gastrin(O.3-3nM) induced significant migration of cells into a denuded area after 6 hours (Control 5.3k1.7; Gastrin InM 38.4k5.4 cells crossing Imm edge in 8 hours p
GENE EXPRESSION
IN INTESTINAL
ADAPTATION
Baksheev, Lyrissa; Dunphy, Jillian L; Taylor, Russell G; Fuller, Peter J Prince Henry’s Institute of Medical Research and Department of Surgery Royal Children’s
Hospital, Australia
Massive small bowel resection (MSBR) is a well characterised model for studying intestinal adaptation. In this model 70%-80% of the small bowel is surgically resected and the remaining bowel re-anastomosed. The residual ileum undergoes a hypertrophic/hyperplastic response; a response is also seen in the jejunum. Our group has previously characterised the response of a number of genes including the proglucagon gene to MSBR. A product of this gene, GLP-2 has recently been shown to be a potent growth factor for the intestinal epithelium via a specific receptor, GLP-2R. The homeobox gene Cdx-2 is thought to regulate proglucagon expression in the bowel. Both Cdx-2 and Cdx-1 are also thought to be involved in the control of epithelial proliferation. The aims of this study are to investigate the effect of MSBR on GLP-2R and Cdx-2 gene expression; and to identify novel genes whose expression is regulated post-MSBR. RNA is isolated using the GTC/CsCI method from whole segments of terminal ileum or from isolated crypt cells obtained from rats pre and post-MSBR. GLP-2R expression is measured by RT-PCR with Southern blot analysis of the PCR products. cDNA probes for rat Cdx-1 and Cdx-2 have been cloned by RT-PCR and used for Northern blot analysis of Cdx gene expression. Differential display-PCR (DD-PCR) is employed to identify novel transcripts. The GLP-2R has been cloned using RT-PCR and the mRNA levels post-MSBR are currently being determined. Cdx-1 and Cdx-2 mRNA levels increase and decrease respectively in response to MSBR. Both Cdx-1 and Cdx-2 mRNA levels are most abundant in the crypts with decreasing levels toward the villus tip. DD-PCR has revealed a number of putative differentially regulated genes which are currently being characterized. Identification and characterization of genes/factors regulating the adaptive response may have implications for both our understanding of and the future treatment of small bowel disease.
STIMULATES
MIGRATION
OF AGS GASTRIC
EPITHELIAL
CELLS
Noble, Peter-John Mantyla; Dockray, Graham J; Varro, Andrea Physiological Laboratory, University of Liverpool, United Kingdom Gastric epithelial cells migrate to form gastric tubules, during epithelial restitution and during tumour invasion. Gastrin regulates the organisation of the gastric epithelium and here we report it stimulates migration of AGS gastric epithelial cells stably transfected with the gastrin-CCKB receptor (AGS-Gr). AGS-Gr cell migration was quantified in a wound healing assay on confluent monolayers and using time lapse videomicroscopy. Cell-cell interactions were investigated by co-culturing AGS-Gr cells with AGS cells lacking the gastrin receptor but constitutively expressing green fluorescent protein (AGS-GFP cells). Gastrin(O.3-3nM) induced significant migration of cells into a denuded area after 6 hours (Control 5.3k1.7; Gastrin InM 38.4k5.4 cells crossing Imm edge in 8 hours p
GENE EXPRESSION
IN INTESTINAL
ADAPTATION
Baksheev, Lyrissa; Dunphy, Jillian L; Taylor, Russell G; Fuller, Peter J Prince Henry’s Institute of Medical Research and Department of Surgery Royal Children’s
Hospital, Australia
Massive small bowel resection (MSBR) is a well characterised model for studying intestinal adaptation. In this model 70%-80% of the small bowel is surgically resected and the remaining bowel re-anastomosed. The residual ileum undergoes a hypertrophic/hyperplastic response; a response is also seen in the jejunum. Our group has previously characterised the response of a number of genes including the proglucagon gene to MSBR. A product of this gene, GLP-2 has recently been shown to be a potent growth factor for the intestinal epithelium via a specific receptor, GLP-2R. The homeobox gene Cdx-2 is thought to regulate proglucagon expression in the bowel. Both Cdx-2 and Cdx-1 are also thought to be involved in the control of epithelial proliferation. The aims of this study are to investigate the effect of MSBR on GLP-2R and Cdx-2 gene expression; and to identify novel genes whose expression is regulated post-MSBR. RNA is isolated using the GTC/CsCI method from whole segments of terminal ileum or from isolated crypt cells obtained from rats pre and post-MSBR. GLP-2R expression is measured by RT-PCR with Southern blot analysis of the PCR products. cDNA probes for rat Cdx-1 and Cdx-2 have been cloned by RT-PCR and used for Northern blot analysis of Cdx gene expression. Differential display-PCR (DD-PCR) is employed to identify novel transcripts. The GLP-2R has been cloned using RT-PCR and the mRNA levels post-MSBR are currently being determined. Cdx-1 and Cdx-2 mRNA levels increase and decrease respectively in response to MSBR. Both Cdx-1 and Cdx-2 mRNA levels are most abundant in the crypts with decreasing levels toward the villus tip. DD-PCR has revealed a number of putative differentially regulated genes which are currently being characterized. Identification and characterization of genes/factors regulating the adaptive response may have implications for both our understanding of and the future treatment of small bowel disease.