Growth factor effects on apoptosis of rat gastric enterochromaffin-like cells.
Enterochromaffin-like (ECL) cells are histamine-containing endocrine cells in the gastric epithelium that show increased density during chronic atrophic gastritis. The current study determined cell number and apoptosis of isolated rat ECL cells in response to several growth factors. Isolated ECL cells from fundic mucosa (enrichment >90%) were grown in serum-free medium over 2-5 days. Cell number was determined by mitochondrial formazan production; apoptosis was measured by Tdt-mediated dUTP nick end labeling reaction and DNA fragmentation-based enzyme-linked immunosorbent assay. Immunocytochemistry and RT-PCR demonstrated the presence of epidermal growth factor receptor, neuronal growth factor receptor (type 1), and fibroblast growth factor (FGF) receptor (type 1). Gastrin (EC50, approximately 2 pM), transforming growth factor-alpha (TGF alpha; 10-30 ng/ml), and basic FGF (bFGF; 1-10 ng/ml) increased the total number of cultured ECL cells. bFGF augmented the gastrin (1 pM)-induced response. Beta-neuronal growth factor (10 ng/ml) and bFGF (2 ng/ml) decreased the programed death of ECL cells. Interleukin-1beta (100 pg/ml, 24 h) stimulated apoptosis 2- to 3-fold in ECL cells, and simultaneous incubation with TGF alpha (20 ng/ml) or bFGF (2 ng/ml) significantly inhibited this effect. ECL cells express specific receptors for gastrin, epidermal growth factor, neuronal growth factor, and FGF. bFGF prolonged ECL cell survival by inhibiting spontaneous apoptosis. Our data further indicate that TGF alpha and bFGF increase ECL cell number by inhibiting cytokine-induced programed cell death.
Mahr
S
S
Department of Medicine II, Technical University of Munich, Germany.
Neumayer
N
N
Kolb
H J
HJ
Schepp
W
W
Classen
M
M
Prinz
C
C
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002452:Cell Count; D002478:Cells, Cultured; D003470:Culture Media; D016207:Cytokines / Q000494:pharmacology; D019861:Enterochromaffin-like Cells / Q000187:drug effects*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology*; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D017978:Receptors, Growth Factor / Q000032:analysis; D016211:Transforming Growth Factor alpha / Q000096:biosynthesis / Q000494:pharmacology
D003470:Culture Media; D016207:Cytokines; D006133:Growth Substances; D009414:Nerve Growth Factors; D017978:Receptors, Growth Factor; D016211:Transforming Growth Factor alpha; D016222:Fibroblast Growth Factor 2