PMID- 15664395
OWN - NLM
STAT- MEDLINE
DCOM- 20050314
LR  - 20131121
IS  - 0008-6363 (Print)
IS  - 0008-6363 (Linking)
VI  - 65
IP  - 3
DP  - 2005 Feb 15
TI  - Diversity and similarity in signaling events leading to rapid Cox-2 induction by 
      tumor necrosis factor-alpha and phorbol ester in human endothelial cells.
PG  - 683-93
AB  - OBJECTIVE: This study examines whether cyclooxygenase 2 (Cox-2) synthesis in 
      human endothelial cells involves different signaling pathways when induced by the 
      proinflammatory cytokine tumor necrosis factor-alpha (TNF alpha) or by the tumor 
      and angiogenic promoter phorbol ester (PMA). Moreover, the hypothesis that 
      reactive oxygen species (ROS) and an altered redox status within the cell are 
      fundamental steps for Cox-2 synthesis is verified. METHODS: Human endothelial 
      cells isolated from umbilical vein (HUVEC) were exposed to PMA and TNF alpha and 
      Cox-2 protein and mRNA levels were evaluated by Western blot and Real-Time 
      Quantitative Reverse Transcription-PCR analysis. Prostaglandin E(2) (PGE(2)) and 
      6-keto prostaglandin F(1 alpha) (6-keto-PGF(1 alpha)) levels were measured in 
      cell medium as an index of Cox-2 activity. Intracellular ROS formation was 
      detected by flow cytometry in HUVEC loaded with the oxidant-sensitive 
      2',7'-dichlorofluorescein diacetate (DCFH-DA) and by nitroblue tetrazolium (NBT) 
      reduction. Reduced and oxidized glutathione (GSH and GSSG) were measured by HPLC. 
      RESULTS: Data show that TNF alpha and PMA signal for early Cox-2 induction 
      through distinct pathways. PMA-induced Cox-2 expression involves a small 
      GTPase-dependent pathway acting via tyrosine kinase, activation of protein kinase 
      C (PKC) and of the mitogen-activated protein kinase (MAPK) ERK1/2. Conversely, 
      MAPK p38 is critical for Cox-2 induction by TNF alpha. Of interest, intracellular 
      ROS generation and consequent GSH/GSSG ratio reduction represents a common step 
      through which PMA and TNF alpha signal for early Cox-2 induction. In addition, we 
      provide evidence that phosphatidylinositol 3 (PI3)-kinase activation plays a 
      regulatory role for Cox-2 synthesis in HUVEC. CONCLUSION: Cox-2 represents a 
      critical link among vascular homeostasis, inflammatory response, angiogenesis and 
      tumor growth. The finding that two independent pathways and an overlapping 
      upstream event signal for Cox-2 induction in HUVEC may be of relevance to develop 
      strategies aimed at selectively interfering with Cox-2 regulating pathways.
FAU - Eligini, Sonia
AU  - Eligini S
AD  - E. Grossi Paoletti Center, Department of Pharmacological Sciences, University of 
      Milan, Via Balzaretti 9, 20133 Milan, Italy.
FAU - Barbieri, Silvia Stella
AU  - Barbieri SS
FAU - Cavalca, Viviana
AU  - Cavalca V
FAU - Camera, Marina
AU  - Camera M
FAU - Brambilla, Marta
AU  - Brambilla M
FAU - De Franceschi, Michela
AU  - De Franceschi M
FAU - Tremoli, Elena
AU  - Tremoli E
FAU - Colli, Susanna
AU  - Colli S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Cardiovasc Res
JT  - Cardiovascular research
JID - 0077427
RN  - 0 (Membrane Proteins)
RN  - 0 (Reactive Oxygen Species)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - EC 1.14.99.1 (Cyclooxygenase 2)
RN  - EC 1.14.99.1 (PTGS2 protein, human)
RN  - EC 1.14.99.1 (Prostaglandin-Endoperoxide Synthases)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases)
RN  - EC 3.6.1.- (GTP Phosphohydrolases)
RN  - GAN16C9B8O (Glutathione)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
RN  - ULW86O013H (Glutathione Disulfide)
SB  - IM
MH  - Cells, Cultured
MH  - Cyclooxygenase 2
MH  - Endothelial Cells/drug effects/enzymology
MH  - Endothelium, Vascular/*drug effects/enzymology
MH  - Enzyme Activation
MH  - GTP Phosphohydrolases/physiology
MH  - Glutathione/physiology
MH  - Glutathione Disulfide/physiology
MH  - Humans
MH  - Membrane Proteins
MH  - Phosphatidylinositol 3-Kinases/physiology
MH  - Prostaglandin-Endoperoxide Synthases/*biosynthesis
MH  - Protein Kinase C/physiology
MH  - Reactive Oxygen Species/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction/methods
MH  - Signal Transduction/*drug effects
MH  - Tetradecanoylphorbol Acetate/*pharmacology
MH  - Tumor Necrosis Factor-alpha/*pharmacology
MH  - p38 Mitogen-Activated Protein Kinases/physiology
EDAT- 2005/01/25 09:00
MHDA- 2005/03/15 09:00
CRDT- 2005/01/25 09:00
PHST- 2004/05/19 00:00 [received]
PHST- 2004/10/13 00:00 [revised]
PHST- 2004/10/19 00:00 [accepted]
PHST- 2005/01/25 09:00 [pubmed]
PHST- 2005/03/15 09:00 [medline]
PHST- 2005/01/25 09:00 [entrez]
AID - S0008-6363(04)00478-X [pii]
AID - 10.1016/j.cardiores.2004.10.024 [doi]
PST - ppublish
SO  - Cardiovasc Res. 2005 Feb 15;65(3):683-93. doi: 10.1016/j.cardiores.2004.10.024.