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Overexpression of manganous superoxide dismutase (MnSOD) in pulmonary endothelial cells confers resistance to hyperoxia
Author(s) -
LindauShepard Barbara,
Shaffer Jacquelin B.,
Del Vecchio Peter J.
Publication year - 1994
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041610207
Subject(s) - hyperoxia , superoxide dismutase , transfection , microbiology and biotechnology , biology , glutathione peroxidase , cell culture , catalase , oxidative stress , clonogenic assay , chemistry , biochemistry , genetics , organic chemistry , oxygen
Treatment of cells or organisms with agents that increase the expression of MnSOD confers resistance to certain types of oxidative damage. However, since these treatments also affect other cellular systems with antioxidant capabilities, the role of MnSOD remains uncertain. To better determine whether increased MnSOD expression confers increased resistance to oxidant stress, a eukaryotic expression vector harboring a mouse MnSOD cDNA was constructed. Bovine lung microvessel endothelial cells were co‐transfected with the MnSOD expression vector and pSV2‐neo, which contains the neo' gene and provides a dominant selectable marker. Control clones were generated by transfecting the cells with pSV2‐neo alone. Stably transfected cell lines were selected and cell lines overexpressing MnSOD were confirmed by Northern blotting, immunoblot analysis, and activity gels. The activities of copper/zinc superoxide dismutase, catalase, and glutathione peroxidase were examined, and no increase in activity of any of these enzymes was detected. Cells were exposed to hyperoxic challenge by treatment with 95% O 2 and 5% CO 2 for 24 h. Viability was assessed by a clonogenic assay. The cell lines that overexpressed MnSOD showed a twofold increase in survival compared to control cells. These results demonstrate a significant resistance to hyperoxia induced oxidative stress in endothelial cells overexpressing MnSOD. © 1994 Wiley‐Liss, Inc.