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Am J Physiol Gastrointest Liver Physiol 271: G104-G112, 1996;
0193-1857/96 $5.00
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AJP - Gastrointestinal and Liver Physiology, Vol 271, Issue 1 104-G112, Copyright © 1996 by American Physiological Society

ARTICLES

Insulin potentiates mitogenic effect of epidermal growth factor on cultured guinea pig gastric mucous cells

S. Ogihara, M. Yamada, T. Saito, M. Shono and K. Rokutan
Fourth Department of Internal Medicine, Tokyo Medical College, Japan.

Epidermal growth factor (EGF) stimulated proliferation of gastric mucous epithelial cells from guinea pigs in serum-free culture conditions. Western blot analysis with antiphosphotyrosine antibody showed that EGF initiated tyrosine phosphorylation of a 170-kDa protein, and this protein was identical to the EGF receptor. Insulin was not mitogenic, but it potentiated the mitogenic effect of EGF. Tyrosine phosphorylation of additional proteins was not induced by the combined actions of insulin and EGF. Stimulation by EGF and/or insulin did not cause a calcium response. However, when insulin was added to cells pretreated with EGF for > 6 h, it elicited a rapid intracellular calcium concentration rise that was reproducible in both cell suspension and single cell analyses. This calcium response coincided with the translocation of protein kinase C (PKC) from the cytosolic to the particulate fraction. Phorbol 12-myristate 13-acetate also caused the translocation and stimulated proliferation of the cells. These results suggest that the calcium-dependent activation of PKC may participate in the potentiation of the mitogenic effect of EGF by insulin.


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