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Induction and Testing of Hypoxia in Cell Culture
07:01

Induction and Testing of Hypoxia in Cell Culture

Published on: August 12, 2011

Regulation of gadd153 mRNA expression by hypoxia in pulmonary artery smooth muscle cells

R Chen1, K S Harrod, J W Olson

  • 1Department of Medicine, Medical University of South Carolina, Charleston, USA. Ruihua_chen@fmc.com

Research Communications in Molecular Pathology and Pharmacology
|January 5, 2002
PubMed

Insights

Hypoxia increases gadd153 mRNA in pulmonary artery smooth muscle cells (PASMCs). This induction is mediated by common signaling pathways, not oxidative stress.

Area of Science:

  • Cell Biology
  • Physiology
  • Molecular Biology

Background:

  • Hypoxia is a known cause of pulmonary hypertension.
  • Pulmonary artery smooth muscle cells (PASMCs) generate oxygen radicals under hypoxic conditions.
  • Oxidative stress is implicated in regulating gadd153 gene expression.

Purpose of the Study:

  • To investigate the effect of hypoxia on gadd153 mRNA expression in PASMCs.
  • To elucidate the signaling pathways involved in hypoxic induction of gadd153 mRNA.

Main Methods:

  • PASMCs were cultured under hypoxic conditions (1% oxygen) for 24 hours.
  • The impact of protein synthesis inhibition, calcium channel blockade (verapamil), G protein inhibition (pertussis toxin), and protein kinase C (PKC) down-regulation was assessed.
  • The effect of antioxidants on gadd153 expression was also examined.

Main Results:

  • Hypoxia significantly increased gadd153 mRNA levels in PASMCs.
  • This induction was independent of protein synthesis.
  • Inhibition of calcium channels, G proteins, and PKC suppressed the hypoxic induction of gadd153 mRNA.
  • Antioxidants did not affect gadd153 expression levels.

Conclusions:

  • Hypoxia induces gadd153 gene expression in PASMCs.
  • The induction of gadd153 by hypoxia involves common signaling pathways, including those regulated by calcium, G proteins, and PKC.
  • Oxidative stress is not the primary mediator of hypoxic gadd153 induction in PASMCs.

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