Effect of transfection with a superoxide dismutase expression plasmid on xanthine/xanthine oxidase-induced cytotoxicity in cultured rat lung cells.
We inserted human Cu, Zn-superoxide dismutase (hSOD) cDNA into the eukaryotic expression plasmid (pRc/CMV) under the control of the cytomegalovirus promoter. The hSOD expression plasmid (pRc/CMV-SOD) was transfected in L2 cells by mean of lipofection. The intracellular SOD activity in pRc/CMV-SOD transfected cells (CMV-SOD cells) was about twice that in host cells. However the level of extracellular SOD activity was similar in CMV-SOD and host cells. When exposed to xanthine (X)/xanthine oxidase (XO) to generate active oxygen species, significantly more CMV-SOD cells than host cells survived. The production of lipid peroxidation in host cells significantly increased in the presence of X/XO, but that in CMV-SOD cells did not change. Thus, transfection with SOD gene effectively prevented X/XO-induced cytotoxicity. The results indicated that increasing the level of intracellular SOD activity protected cells against extracellular superoxide anion stress.
Komada
F
F
Department of Hospital Pharmacy, School of Medicine, Kobe University, Japan.
Nishiguchi
K
K
Tanigawara
Y
Y
Wu
X Y
XY
Iwakawa
S
S
Okumura
K
K
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D015316:Genetic Therapy; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects; D010957:Plasmids; D051381:Rats; D013482:Superoxide Dismutase / Q000235:genetics*; D014162:Transfection; D019820:Xanthine; D014969:Xanthine Oxidase / Q000494:pharmacology*; D014970:Xanthines / Q000494:pharmacology*
D014970:Xanthines; D019820:Xanthine; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase