The mitogenic inhibitory effect of phorbol esters is associated with decreased activation of phosphatidylinositol-3 kinase. Weiss, Robert H., and Ann P. Yabes. Division of Nephrology, Department of Internal Medicine, University of California, Davis, CA. 95616 and Department of Veteran's Affairs Northern California System of Clinics, Pleasant Hill, CA. 94523
APStracts 2:0320C, 1995.
In contrast to their role as potent tumor promoters, phorbol esters can cause inhibition of cell growth. Since the effect of phorbol esters occurs through activation of protein kinase C (PKC), and because activated PKC is translocated to the membrane placing it in a position to act on the intracellular portion of the growth factor receptor, we asked whether this inhibitory effect is mediated through the action of phorbol-12-myristate-13-acetate (PMA) on receptor association with the signal transfer proteins. When added to rat vascular smooth muscle (VSM) cells concurrently with bFGF, PMA at 100 ng/ml completely inhibits bFGF-stimulated DNA synthesis. Under the same growth-inhibitory conditions of PMA addition, aggregation of phosphatidylinositol-3 kinase (PI3K) to the FGF receptor and tyrosine phosphorylation of the 85 kD regulatory component of the signal transfer protein PI3K are reduced by 94% and 79%, respectively. PI3K catalytic activity, as measured by conversion of phosphatidylinositol to phosphatidylinositol monophosphate, is decreased 88% by PMA addition. This effect is not specific to PI3K, as aggregation of PLC -[gamma]1 to the activated bFGF receptor is also decreased by PMA treatment. In addition, the PI3K inhibitor wortmannin markedly attenuates bFGF-stimulated VSM cell growth in a dose-dependent manner. These data suggest that the site of growth inhibition by PMA in vascular smooth muscle cells lies upstream of signal transfer particle aggregation and that such growth arrest may be mediated through inhibition of activation of PI3K.

Received 5 July 1995; accepted in final form 28 August 1995.
APS Manuscript Number C402-5.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 23 September 1995.