Hypoxic modulation of manganese superoxide dismutase promoter activity and gene expression in lung epithelial cells

Tauni Ohman, Gregory Parish, Robert Jackson

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

We investigated the effects of hypoxia (< 2.5% O2) on rat manganese superoxide dismutase (MnSOD) gene promoter-luciferase reporter constructs in transiently transfected lung epithelial cells (A549, 1,2, and E1A-T2) and fibroblasts (R9Ab). We cloned MnSOD promoter-luciferase reporter constructs (numbers refer to length in base pairs |bp| in the 5′ direction from the transcription initiation site): 2,505, 1,064, 507, 405, and 289 into pGL2-Basic, a promoterless, firefly luciferase vector. Lung cells were transfected with MnSOD promoter-reporter constructs with or without thymidine kinase-driven Renilla luciferase (pRL-TK), and were exposed to air/5% CO2 or hypoxia (2.5% O2/5% CO2/balance N2) for 24 h. Hypoxia caused a significant (by two-way analysis of variance) consistent increase in luciferase in the A549 cell (human lung carcinoma) line. Greatest expression (> 3-fold increase) in hypoxia was associated with the 2,505-bp MnSOD promoter (normalized to cellular protein). Azide (10 μM) did not increase expression of the MnSOD reporter constructs. The 289-bp promoter was sufficient to express the reporter in air and to increase its expression in hypoxia. Promoter activity of the rat MnSOD 5′ region, assessed by luciferase reporter constructs in A549 cells, increased in hypoxia. The increase was exclusive to A549 cells and did not occur in other cells.

Original languageEnglish
Pages (from-to)119-127
Number of pages9
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume21
Issue number1
StatePublished - Dec 1 1999
Externally publishedYes

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Azides
Luciferases
Gene expression
Superoxide Dismutase
Rats
Epithelial Cells
Modulation
Gene Expression
Lung
Air
Proteins
Hypoxia
A549 Cells

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

Cite this

Hypoxic modulation of manganese superoxide dismutase promoter activity and gene expression in lung epithelial cells. / Ohman, Tauni; Parish, Gregory; Jackson, Robert.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 21, No. 1, 01.12.1999, p. 119-127.

Research output: Contribution to journalArticle

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