Epidermal growth factor stimulates phospholipase D independent of phospholipase C, protein kinase C or phosphatidylinositol-3 kinase activation in immortalized rabbit corneal epithelial cells

Current Eye Research
Y Zhang, R A Akhtar

Abstract

Activation of phospholipase D (PLD) is believed to be an important signaling pathway involved in cell growth and differentiation in several tissues, in response to a variety of mitogens. The aim of the present study was to investigate the effect of epidermal growth factor (EGF) on PLD activity in rabbit corneal epithelial cells (RCEC). We have also examined whether the EGF effect is dependent on concurrent activation of phospholipase C (PLC), protein kinase C (PKC) or phosphatidylinositol 3-kinase (PI-3-kinase) in these cells. RCEC, immortalized with adenovirus SV-40, were cultured until they became confluent. The cells were labeled with [3H]myristic acid and incubated with or without EGF or other agents for specified time intervals. PLD activity was measured by quantifying [3H]phosphatidylethanol in cells incubated in the presence of ethanol. PLC activity was determined by measuring the radioactivity in inositol trisphosphate in myo[3H]inositol-labeled RCEC. PI 3-kinase activity was assessed by measuring the production of PIP3 in 32P-labeled cells. Addition of EGF to RCEC stimulated PLD activity in a time- and dose-dependent manner. The maximal effect was observed with 150 ng/ml EGF and at 10 min of incubation. The PLD activit...Continue Reading

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Citations

May 5, 2009·Biochimica Et Biophysica Acta·Chang Sup LeeSung Ho Ryu
Nov 26, 2015·Trends in Molecular Medicine·Agnes LimLouis Tong
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