Insulin potentiates mitogenic effect of epidermal growth factor on cultured gastric mucous cells. Ogihara, Shoji, Masahiko Yamada, Toshihiko Saito, Masayuki Shono, and Kazuhito Rokutan. Fourth Department of Internal Medicine, Tokyo Medical College, Tokyo160, Japan; and Department of Nutrition, School of Medicine, The University of Tokushima. Tokushima 770, Japan
APStracts 3:0013G, 1996.
Epidermal growth factor (EGF) stimulated proliferation of gastric mucous epithelial cells from guinea pigs in serum-free culture conditions. Western blot analysis with anti-phosphotyrosine antibody showed that EGF initiated tyrosine phosphorylation of a 170 kilodalton 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 more than 6 h, it elicited a rapid [Ca2+]i rise, which 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.

Received 10 August 1993; accepted in final form 26 December 1995.
APS Manuscript Number G317-3.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 22 January 96