Role of stimulation of arachidonic acid release in the proliferative response of 3T3 mouse fibroblasts to platelet-derived growth factor.

Abstract

Human platelet-derived growth factor (PDGF) stimulates release of arachidonic acid from cellular phospholipids, synthesis and release of prostaglandins from the cell, and initiation of DNA synthesis in cultures of 3T3 Swiss mouse fibroblasts at similar concentrations with four independent preparations representing a million-fold range of purification. Stimulation of arachidonic acid and prostaglandin release is an early event (beginning within minutes) in the response to PDGF treatment. Incubating cells with PDGF at 4 degrees C followed by washing leads to activation of arachidonic acid release on warming the cells to 37 degrees C, consistent with binding of the factor to the cell surface. PDGF-stimulated arachidonic acid release, prostaglandin release, and initiation of DNA synthesis are all inhibited by phenylglyoxal at similar concentrations. These results suggest that activation of arachidonic acid release from phospholipids plays an essential role in the mechanism by which PDGF-stimulates the initiation of DNA synthesis in 3T3 cells. The stimulation of initiation of DNA synthesis by PDGF does not appear to be mediated by the synthesis of prostaglandins or other known arachidonic acid metabolites because neither indomethacin (a fatty acid cyclooxygenase inhibitor) nor phenidone (a lipoxygenase inhibitor) inhibit initiation of DNA synthesis at concentrations which inhibit arachidonic acid metabolism. Although the activation of arachidonic acid release by PDGF is a calcium-dependent process, a simple calcium flux appears unimportant to the mechanism of activation. Evidence was also obtained against an involvement of sodium fluxes or proteolytic activity in the mechanism of stimulating arachidonic acid release by PDGF or serum.