CuZn-SOD suppresses the bovine papillomavirus-induced proliferation of fibroblasts.

Eukaryotic cells continuously produce reactive oxygen species (ROS) and have mechanisms to control ROS levels. ROS have been shown to mediate cell proliferation and transformation. We studied the effect of CuZn-superoxide dismutase (CuZnSOD) on the focus-forming ability of bovine papillomavirus (BPV-1) wtDNA and hypertransforming mutant of its major oncoprotein E5, E5-17S. We found that CuZnSOD suppresses the focus-forming ability of BPV-1 wtDNA and E5 oncoprotein. Significantly fewer foci were detected in pCGCuZnSOD- and BPV-1 DNA-cotransfected cell culture compare to BPV-1 DNA-transfected cell culture (p<0.001). CuZnSOD decreases the rate of cell proliferation in both non-transformed C127 and BPV-1- and E5-transformed cell lines. CuZnSOD decelerates cell entry into the S phase of the cell cycle and has a suppressing effect on the actively dividing cells. As the transformed cells proliferate faster than normal cells when confluent, CuZnSOD inhibits the growth of foci. These results indicate that superoxide radicals may be involved in signaling for cell proliferation and that SOD suppresses cell proliferation.

Bovine papillomavirus type 1 oncoprotein E5 stimulates the utilization of superoxide radicals in the mouse fibroblast cell line C127.

The major transforming protein of bovine papillomavirus type 1 (BPV-1) is a small hydrophobic polypeptide, the E5 gene product, localized in the cellular membranes and modulating various pathways in the cell. Many studies have shown that reactive oxygen species (ROS) are essential in several biological processes, including cell transformation by oncogenes, but unregulated ROS are highly toxic to cells. We studied the effect of the bovine papillomavirus protein E5 and its mutants on the level of the superoxide radicals in the mouse fibroblast cell line C127. The superoxide level in C127 cells transfected with the E5-expressing plasmids were measured by nitroblue tetrazolium reduction. Relative concentrations of intracellular peroxide were determined by using 2,7-dichlorofluorescin diacetate. Our results showed that all transforming mutants of E5 reduced the level of superoxide in C127 cells, besides the activity of superoxide dismutase (SOD) and level of peroxides was not altered. In the presence of neopterin, an inhibitor of the superoxide-producing enzymes, the reduction of superoxide level correlated with the transforming ability of the E5-mutants. The inhibitor of the protein tyrosine kinase, tyrphostin 25 and inhibitors of oxygenases of the arachidonic acid metabolism, aspirin and nordihydroguaiaretic acid, blocked the effect of BPV-1 E5. We conclude that BPV-1 E5 and its transforming mutants are able to modulate the level of superoxide and stimulate the utilization of superoxide through protein tyrosine kinases and oxygenases of the arachidonic acid metabolism.

Two antioxidative lactobacilli strains as promising probiotics.

Two antioxidative strains tentatively identified as Lactobacillus fermentum, E-3 and E-18, were isolated from intestinal microflora of a healthy child. Survival time of these strains in the presence of reactive oxygen species (ROS), like hydrogen peroxide, superoxide anions and hydroxyl radicals, was significantly increased compared with a non-antioxidative strain, and also was quite similar to a highly ROS resistant strain of Salmonella typhimurium. E-3 and E-8 contain a remarkable level of glutathione, express Mn-SOD, which is important for the prevention of lipid peroxidation, and secrete hydrogen peroxide. Their significant antimicrobial activity combined with antioxidative properties may serve as defensive principles in the intestinal microbial ecosystem and overcome exo- and endogenous oxidative stress.