Cytosolic phospholipase A2 mediates arachidonoyl phospholipid hydrolysis in immortalized rat brain endothelial cells stimulated by oxidized LDL

Endothelial cell-pericyte cocultures induce PLA2 protein expression through activation of PKCα and the MAPK/ERK cascade

“…In agreement with our previous results in ECs (33), the intense staining of cPLA 2 seems to be associated with the nuclear compartment (Fig. 7).…”

“…The cell line was already characterized, and our cell cultures were prepared and characterized according to previously described procedures (31,32). Morphological changes and cell viability determined by [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-tetrazolium bromide] test were assessed as reported previously (29,33). Pure microvessel pericyte cultures were prepared from bovine retinas as described previously (34).…”

“…After a coincubation for 24 h in EC/pericyte medium (50% DMEM plus F10 Ham's, each containing 10% FBS), inserts containing conditioning cells were discarded, whereas cells on the plate were washed three times with ice-cold PBS and scraped with a rubber policeman. Lysates were performed as described (29,33). EC or pericyte (40,000 cells/cm 2 ) monocultures were established in parallel as controls.…”

“…Then, it was acidified with 1 M HCl to bring the pH to 3.0 (medium turns yellow) and frozen for subsequent analysis. Acidified medium was extracted three times in equal volumes of ethyl acetate (33). The organic layer was evaporated under nitrogen, and the residue was dissolved in ethanol.…”

Melanoma-Induced Endothelial Cell Growth Involves Phospholipase A2 and COX2 Upregulation

“…Phospholipases A 2 are a diverse group of enzymes that catalyze the hydrolysis of the sn-2 substituent from glycerophospholipid substrates to yield a free fatty acid and 2-lysophospholipid acceptors (Balsinde et al, 2002). They are the rate limiting step for the AA production from membrane phospholipids, which in turn is the major precursor to prostaglandins, leukotrienes, and hydroxyeicosatetraenoic acids (via the cyclooxygenase, lipoxygenase, and epoxygenase pathways, respectively) that increase cell proliferation in response to various agonists in different cell types (Anfuso et al, 2007;Balsinde et al, 2002;Chakraborti, 2003;Lupo et al, 2002). Among the PLA 2 s is an 85 kDa cPLA 2 that requires Ca 2+ for catalysis, and a calcium independent PLA 2 , iPLA 2 β, for which several potential functions have been proposed, including a housekeeping role in phospholipid remodeling and a signaling role in cell growth, apoptosis, secretion, inflammation, and oxidant-induced cell injury (Chakraborti, 2003).…”