Cytosolic Phospholipase A2-Mediated ICAM-1 Expression Is Calcium Dependent

Author: Barnett C.C.   Moore E.E.   Silliman C.C.   Abdalla E.K.   Partrick D.A.   Curley S.A.  

Publisher: Academic Press

ISSN: 0022-4804

Source: Journal of Surgical Research, Vol.99, Iss.2, 2001-08, pp. : 307-310

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Abstract

Background. Some human malignancies such as virus-related hepatocellular cancer arise in a setting of chronic inflammation. Upregulation of ICAM-1 is a seminal late event in malignant transformation following chronic inflammation. Cytosolic phospholipase A2 (cPLA2) is a lipid-mediator activated by inflammatory stimuli, which has been shown to mediate ICAM-1 upregulation. As lipid mediators are known to work via calcium-dependent mechanisms in nearly all mammalian cells, we hypothesize that inflammatory-mediated ICAM-1 upregulation is dependent on both cPLA2 and intracellular calcium.Materials and methods. HUVEC were chosen as a representative cell line as they emulate hepatic sinusoids and are a well-established cell model. These were grown to confluence in T-25 flasks and stimulated with TNF-α or LPS for 6 h. Additional groups were preincubated with AACOCF3 (a specific cPLA2 inhibitor) or BAPTA A.M. (a specific inhibitor of intracellular Ca2+) prior to being exposed to inflammatory stimuli. ICAM-1 expression was determined by mean fluorescent intensity (MFI) as measured by FITC-labeled moAb to ICAM-1 via FACS. The role of intracellular Ca2+ on cPLA2 activity was determined by thin-layer chromatography. Groups were compared using ANOVA with Scheffe's post hoc analysis; *P < 0.05 vs control, †P < 0.05 vs LPS and TNF-α was considered significant; N ≥ 4 all experimental groups.Results. Both cPLA2 and Ca2+ inhibition significantly inhibited inflammatory upregulation of ICAM-1. Pretreatment with BAPTA A.M. attenuated HUVEC cPLA2 activity in response to LPS. These findings suggest that appropriate molecular target suppression may prevent malignant degeneration in the presence of chronic inflammation.

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