Gastrin activates nuclear factor kappaB (NFkappaB) through a protein kinase C dependent pathway involving NFkappaB inducing kinase, inhibitor kappaB (IkappaB) kinase, and tumour necrosis factor receptor associated factor 6 (TRAF6) in MKN-28 cells transfected with gastrin receptor.
BACKGROUND
We previously reported that gastrin induces expression of CXC chemokines through activation of nuclear factor kappaB (NFkappaB) in gastric epithelial cells that express gastrin receptor.


OBJECTIVE
To clarify gastrin receptor mediated signals leading to activation of NFkappaB.


METHODS
MKGR26 cells were created by transfecting gastrin receptor cDNA into MKN-28 cells. Degradation of inhibitor kappaB (IkappaB) and phosphorylation of protein kinase C (PKC)-delta were both detected by western blot analysis. NFkappaB activation was determined by luciferase assay and electrophoretic mobility shift analysis.


RESULTS
Gastrin induced degradation of IkappaB-alpha and activation of NFkappaB, which was abolished by the selective gastrin receptor antagonist L-740,093 and the general PKC inhibitor GF109203X. Gastrin induced phosphorylation of PKC-delta, and its inhibitor rottlerin partially suppressed NFkappaB activation. However, the mitogen activated protein kinase (MAPK) kinase inhibitor PD98059, p38 MAPK inhibitor SB203580, and tyrphostin AG1478 had no effect on NFkappaB activation. Introduction of the dominant negative mutant of IkappaB kinase, of NFkappaB inducing kinase, and of tumour necrosis factor receptor associated factor 6 (TRAF6), but not that of TRAF2, inhibited gastrin induced activation of NFkappaB.


CONCLUSIONS
Gastrin activates NFkappaB via a PKC dependent pathway which involves IkappaB kinase, NFkappaB inducing kinase, and TRAF6.
Department of Internal Medicine and Molecular Science, Graduate School of Medicine, Osaka University, Japan.
M
M
Ogasa
Y
Y
Miyazaki
S
S
Hiraoka
S
S
Kitamura
Y
Y
Nagasawa
O
O
Kishida
T
T
Miyazaki
T
T
Kiyohara
Y
Y
Shinomura
Y
Y
Matsuzawa
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D005753:Gastric Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005755:Gastrins / Q000494:pharmacology*; D006168:Guinea Pigs; D006801:Humans; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D011506:Proteins / Q000502:physiology*; D011949:Receptors, Cholecystokinin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects; D048029:TNF Receptor-Associated Factor 6; D014162:Transfection; D014407:Tumor Cells, Cultured; D000097782:NF-kappaB-Inducing Kinase
D018076:DNA, Complementary; D005755:Gastrins; D016328:NF-kappa B; D011493:Protein Kinase C; D017346:Protein Serine-Threonine Kinases; D011506:Proteins; D011949:Receptors, Cholecystokinin; D048029:TNF Receptor-Associated Factor 6; D000097782:NF-kappaB-Inducing Kinase