INDUCTION OF IN-VITRO ANGIOGENESIS IN THE ENDOTHELIAL-DERIVED CELL-LINE, EA HY926, BY ETHANOL IS MEDIATED THROUGH PKC AND MAPK

We have previously shown that ethanol-induced injury to the gastric mu cosa triggers increased expression of the angiogenic factors, basic fi broblast growth factor (bFGF) and vascular endothelial growth factor ( VEGF) and angiogenesis. To further investigate ethanol-induced angioge nesis, we used an in vitro angiogenesis model which employs the abilit y of an endothelial-derived cell line (EA hy926) to form tubelike stru ctures resembling capillaries when plated on the matrix material, Matr igel. We report that serum-starved EA hy926 cells, incubated for as li ttle as 5 minutes with ethanol concentrations of 1.0-2.5%, formed tube like structures reflecting in vitro angiogenesis. Control cells, not i ncubated with ethanol, did not form tubelike structures. incubation fo r 5 minutes with 2.5% ethanol resulted in increased activities of PKC and MAP kinase (ERK2) by 1.6-fold (p < 0.05) and 2.3-fold (P < 0.001), respectively. Futhermore, inhibitors of the MAPK kinase, MEK (PD98059 ) and PRC (GF 109203X) prevented the induction of in vitro angiogenesi s by ethanol, (C) 1998 Academic Press.