Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 380, Issue 2, pp 193–204

NADPH oxidase isoform selective regulation of endothelial cell proliferation and survival

  • Hitesh Peshavariya
  • Gregory J. Dusting
  • Fan Jiang
  • Lesley R. Halmos
  • Christopher G. Sobey
  • Grant R. Drummond
  • Stavros Selemidis
Original Article

DOI: 10.1007/s00210-009-0413-0

Cite this article as:
Peshavariya, H., Dusting, G.J., Jiang, F. et al. Naunyn-Schmied Arch Pharmacol (2009) 380: 193. doi:10.1007/s00210-009-0413-0

Abstract

Proliferation and apoptosis of endothelial cells are crucial angiogenic processes that contribute to carcinogenesis and tumor progression. Emerging evidence implicates the regulation of proliferation and apoptosis by reactive oxygen species (ROS) such as superoxide and hydrogen peroxide (H2O2). In the present study, we investigated the roles of the ROS-generating Nox4- and Nox2-containing reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidases in proliferation of human endothelial cells by examining the impact of these enzyme systems on (1) specific proliferative and tumorigenic kinases, extracellular regulated kinase1/2 (ERK1/2) and Akt, (2) cytoskeletal organization, and (3) the mechanisms that influence cellular apoptosis. ROS production and the expression of NADPH oxidase subunit Nox4, but not Nox2, were markedly higher in proliferating than in quiescent endothelial cells. Addition of the H2O2 scavenger catalase or downregulation of Nox4 protein with specific siRNA reduced ROS levels, cell proliferation, and ERK1/2 phosphorylation but had no effect on either cell morphology or caspase 3/7 activity. Although downregulation of Nox2 protein with siRNA also reduced ROS production and cell proliferation, it caused an increase in caspase 3/7 activity, reduced Akt phosphorylation, and caused cytoskeletal disorganization. Therefore, in endothelial cells, Nox4-derived H2O2 activates ERK1/2 to promote proliferation, whereas Nox2-containing NADPH oxidase maintains the cytoskeleton and prevents apoptosis to support cell survival. Our study provides a new understanding of the molecular mechanisms that underpin endothelial cell survival and a rationale for the combined suppression of Nox4- and Nox2-containing NADPH oxidases for unwanted angiogenesis in cancer.

Keywords

Cancer Proliferation Apoptosis Endothelial cells Superoxide NADPH oxidase 

Abbreviations

DPI

Diphenylene iodonium

FAD

Flavin adenine dinucleotide

FBS

Fetal bovine serum

H2-DCFDA

Dichlorodihydrofluorescein-diacetate

HMECs

Human microvascular endothelial cells

HUVEC

Human umbilical vein endothelial cells

H2O2

Hydrogen peroxide

ERK1/2

Extracellular regulated kinase1/2

l-NAME

NG-Nitro-l-arginine methyl ester

170-ODYA

17-Octadecynoic acid

ROS

Reactive oxygen species

PI-3 K

Phosphatidyl inositol 3-kinase

TNF-α

Tumor necrosis factor-alpha

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Hitesh Peshavariya
    • 1
  • Gregory J. Dusting
    • 1
  • Fan Jiang
    • 1
  • Lesley R. Halmos
    • 2
  • Christopher G. Sobey
    • 2
  • Grant R. Drummond
    • 2
  • Stavros Selemidis
    • 2
  1. 1.Bernard O’Brien Institute of MicrosurgeryThe University of MelbourneFitzroyAustralia
  2. 2.Department of PharmacologyMonash UniversityClaytonAustralia

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