COOH-terminal 26-amino acid residues of progastrin are sufficient for stimulation of mitosis in murine colonic epithelium in vivo.
Transgenic mice (hGAS) that overexpress human progastrin are more susceptible than wild-type mice (FVB/N) to the induction of colonic aberrant crypt foci (ACF) and adenomas by the chemical carcinogen azoxymethane. We have previously shown significantly increased levels of colonic mitosis in hGAS compared with FVB/N mice after gamma-radiation. To investigate whether the effects of progastrin observed in hGAS colon require the presence of other forms of circulating gastrin, we have crossed hGAS (hg(+/+)) with gastrin knockout (G(-/-)) mice to generate mice that express progastrin and no murine gastrin (G(-/-)hg(+/+)). After azoxymethane, G(-/-)hg(+/+) mice developed significantly more ACF than control G(-/-)hg(-/-) mice (which do not express any forms of gastrin). G(-/-)hg(+/+) mice also exhibited significantly increased colonic mitosis both before and after exposure to 8 Gray Gy gamma-radiation or 50 mg/kg azoxymethane compared with G(-/-)hg(-/-). Treatment of G(-/-)hg(-/-) mice with synthetic progastrin (residues 21-101 of human preprogastrin) or G17 extended at its COOH terminus corresponding to the COOH-terminal 26-amino-acid residues of human preprogastrin (residues 76-101, G17-CFP) resulted in continued colonic epithelial mitosis after gamma-radiation, whereas glycine-extended gastrin-17 and the COOH-terminal tryptic fragment of progastrin [human preprogastrin-(96-101)] had no effect. Immunoneutralization with an antibody against G17-CFP before gamma-radiation significantly decreased colonic mitosis in G(-/-)hg(+/+) mice to levels similar to G(-/-)hg(-/-). We conclude that progastrin does not require the presence of other forms of gastrin to exert proliferative effects on colonic epithelia and that the portion of the peptide responsible for these effects is contained within amino acid residues 76-101 of human preprogastrin.
Deptartment of Medicine, 5th Fl. UCD Bldg., Daulby St., Liverpool L69 3BX, UK.
P D
PD
Ottewell
A
A
Varro
G J
GJ
Dockray
C M
CM
Kirton
A J M
AJ
Watson
T C
TC
Wang
R
R
Dimaline
D M
DM
Pritchard
D000595:Amino Acid Sequence; D000818:Animals; D000963:Antimetabolites; D017209:Apoptosis / Q000187:drug effects; D001397:Azoxymethane / Q000494:pharmacology; D001973:Bromodeoxyuridine; D002273:Carcinogens / Q000494:pharmacology; D003106:Colon / Q000166:cytology* / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects; D004847:Epithelial Cells / Q000187:drug effects*; D005720:Gamma Rays; D005755:Gastrins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D005838:Genotype; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008934:Mitogens; D008938:Mitosis / Q000187:drug effects*; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D011498:Protein Precursors / Q000737:chemistry / Q000494:pharmacology*
D000963:Antimetabolites; D002273:Carcinogens; D005755:Gastrins; D008934:Mitogens; D010446:Peptide Fragments; D011498:Protein Precursors; C013706:big gastrin; D001973:Bromodeoxyuridine; D001397:Azoxymethane