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Secretory Phospholipase A2 Inhibition Attenuates Intercellular Adhesion Molecule-1 Expression in Human Esophageal Adenocarcinoma Cells  Miral R. Sadaria,

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Presentation on theme: "Secretory Phospholipase A2 Inhibition Attenuates Intercellular Adhesion Molecule-1 Expression in Human Esophageal Adenocarcinoma Cells  Miral R. Sadaria,"— Presentation transcript:

1 Secretory Phospholipase A2 Inhibition Attenuates Intercellular Adhesion Molecule-1 Expression in Human Esophageal Adenocarcinoma Cells  Miral R. Sadaria, MD, Xianzhong Meng, MD, PhD, David A. Fullerton, MD, T. Brett Reece, MD, Roopali R. Shah, BA, Frederick L. Grover, MD, Michael J. Weyant, MD  The Annals of Thoracic Surgery  Volume 91, Issue 5, Pages (May 2011) DOI: /j.athoracsur Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

2 Fig 1 The ICAM-1 expression in human esophageal adenocarcinoma cells (FLO-1) stimulated with TNF-α. Stimulating FLO-1 cells with 20 ng/mL of TNF-α increased total cellular (immunoblotting), cell-surface (flow cytometry), and secreted (ELISA) ICAM-1 expression 2.6-fold, 2.0-fold, and 2.3-fold, respectively, compared with vehicle control. (*p < 0.05 compared with vehicle control, immunoblotting n = 5, flow cytometry n = 7, ELISA n = 6.) (ICAM-1 = intercellular adhesion molecule-1; ELISA = enzyme-linked immunosorbent assay; TNF-α = tumor necrosis factor-alpha.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

3 Fig 2 Total cellular ICAM-1 expression attenuated by inhibition of group IIa sPLA2 in human esophageal adenocarcinoma cells (FLO-1). A representative Western blot demonstrates a dose-dependent attenuation of total cellular ICAM-1 expression in FLO-1 cells when treated with specific group IIa sPLA2 inhibitor (A). A density ratio was created by dividing the density of the ICAM-1 by the density of the GAPDH band. The density ratio of each sample was then divided by the vehicle control, giving the vehicle control an adjusted unit value of 1.0. All doses of sPLA2 inhibitor reduced total cellular ICAM-1 expression compared with control (B). (*p < 0.05 compared with vehicle control and 15 μM dose; **p < compared with vehicle control and all doses, n = 5.) (GAPDH = glyceraldehyde-3-phosphate dehydrogenase; ICAM-1 = intercellular adhesion molecule-1; sPLA2 = secretory phospholipase A2.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

4 Fig 3 Cell-surface ICAM-1 expression attenuated by inhibition of group IIa sPLA2 in human esophageal adenocarcinoma cells (FLO-1). A representative flow cytometry histogram demonstrates a reduction in mean intensity fluorescence and, thus, cell-surface ICAM-1 expression after treatment of FLO-1 cells with specific group IIa. sPLA2 inhibitor (A). The mean intensity fluorescence of each sample was normalized to the vehicle control, giving the vehicle control an adjusted unit of 1.0. Cell-surface ICAM-1 expression decreased at the 15 μM dose (B). (*p < compared with vehicle control and all doses, n = 7.) (ICAM-1 = intercellular adhesion molecule-1; PE = phycoerythrin; sPLA2 = secretory phospholipase A2.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

5 Fig 4 Secreted ICAM-1 expression attenuated by inhibition of group IIa sPLA2 in human esophageal adenocarcinoma cells (FLO-1). Specific group IIa sPLA2 inhibitor treatment attenuated ICAM-1 secretion by FLO-1 cells at the 10 μM and 15 μM dose. (*p < 0.05 compared with vehicle control; **p < 0.05 compared with vehicle control and 5 μM dose, n = 6.) (ICAM-1 = intercellular adhesion molecule-1.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

6 Fig 5 NF-κB activation attenuated by inhibition of group IIa sPLA2 in human esophageal adenocarcinoma cells (FLO-1). A representative Western blot demonstrates downregulation of NF-κB activation in FLO-1 cells when treated with specific group IIa sPLA2 inhibitor (A). The phosphorylated and total NF-κB densities were normalized to the glyceraldehyde-3-phosphate dehydrogenase on their respective membranes. The normalized phosphorylated NF-κB density ratio was then divided by the normalized total NF-κB density ratio. This adjusted unit was then normalized to the vehicle control, giving the vehicle control an adjusted unit value of 1.0. A decrease in NF-κB activation was demonstrated specifically at the 15 μM dose of sPLA2 inhibitor (B). (*p < 0.05 compared with vehicle control and 5 μM dose, n = 5.) (NF-κB = nuclear factor-kappa beta). The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

7 Fig 6 Schematic illustrating the proposed relationship between sPLA2 and the TNF-α signaling pathway and the proposed mechanism of action of the specific sPLA2 inhibitor. sPLA2 catalyzes the hydrolysis of membrane phospholipids to generate free fatty acids such as arachidonic acid. These lipid mediators activate protein kinase Cζ, which has been suggested to be involved in TNF-α-mediated NF-κB activation and, ultimately, ICAM-1 expression. The specific sPLA2 inhibitor functions by inhibiting the hydrolysis of membrane phospholipids as denoted by the X in the figure, thus preventing protein kinase Cζ activation and leading to attenuated TNF-α-mediated NF-κB activation and ICAM-1 expression. (ICAM-1 = intercellular adhesion molecule-1; NF-κB = nuclear factor-kappa beta; sPLA2 = secretory phospholipase A2; TNF-α—tumor necrosis factor-alpha.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

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