Expression of human Mn SOD in Chinese hamster ovary cells confers protection from oxidant injury.
Manganese superoxide dismutase (Mn SOD) is an important component of antioxidant defense in aerobic cells because of its location in the mitochondria, a significant source of oxygen radicals and an important target of oxidant injury. To test the hypothesis that increased mitochondrial Mn SOD protects from oxidant injury, Chinese hamster ovary (CHO) cells were transfected with a eukaryotic expression vector containing the human Mn SOD cDNA. In recombinant CHO cells, Mn SOD activity was increased threefold over wild-type controls. Acute survival during paraquat exposure (0-500 microM) was significantly improved in CHO cells expressing human Mn SOD, with 71% of recombinant CHO cells surviving at the 50% lethal dose (LD50) for wild-type CHO controls. Cell growth following exposure to paraquat (100 microM) was also significantly improved in recombinant CHO cells. CHO cells expressing human Mn SOD continued to grow and divide after paraquat exposure, whereas growth of wild-type CHO cells was negligible. Protection against oxidant-induced injury was directly related to increased Mn SOD, occurring in the absence of changes in other antioxidant enzymes including catalase, Cu,Zn SOD, and glutathione associated cellular antioxidant mechanisms. We conclude that increased expression of human Mn SOD in vitro directly confers protection against oxidant injury.
Warner
B
B
Children's Hospital Medical Center, Division of Pulmonary Biology, Cincinnati, Ohio 45229-2899.
Papes
R
R
Heile
M
M
Spitz
D
D
Wispé
J
J
D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D006224:Cricetinae; D004247:DNA; D004351:Drug Resistance; D006801:Humans; D008928:Mitochondria / Q000201:enzymology; D010269:Paraquat / Q000037:antagonists & inhibitors* / Q000494:pharmacology / Q000506:poisoning; D011995:Recombination, Genetic; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D016019:Survival Analysis; D014162:Transfection
D000975:Antioxidants; D004247:DNA; D013482:Superoxide Dismutase; D010269:Paraquat