• Asian Pacific Journal of Cancer Prevention
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• v.13 no.6
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• pp.2571-2576
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• 2012

Breast cancer is the first of the most common ten cancers in Iraq. Its etiology is multifactorial, oxidative stress and lipid peroxidation being suggested to play important roles in carcinogenesis. The purpose of this study was to investigate the oxidant-antioxidant status in breast cancer patients, by measuring SOD isoenzyme activities (total SOD, CuZn-SOD, Mn-SOD and EC-SOD) in plasma and breast tumors, and by estimating thiobarbituric reactive substances (TBRS) in tissue homogenates. General increase in total SOD activity was observed in plasma and tissue samples of breast tumors, greater in the malignant when compared to benign group (p<0.05). Mn-SOD showed a significant decrease in tissue malignant samples (p<0.05), and insignificant decrease in plasma malignant samples compared with control and benign samples. Plasma EC-SOD activity in both patient benign and malignant breast tumors demonstrated 3.5% and 22.8% increase, respectively. However, there was a decrease in tissue EC-SOD activity in malignant breast tumors when compared with benign. A similar tendency was noted for TBRS. We suggest that elevated total SOD might reflect a response to oxidative stress, and then may predict a state of excess reactive oxygen species in the carcinogenesis process. If there is proteolytic removal of the heparin binding domain, EC-SOD will lose its affinity for the extracellular matrix and diffuse out of the tissue. This will result in a decreased EC-SOD activity, thus leading to an increase in the steady-state concentration of $O^{2-}$ in this domain, and increase in EC-SOD activity in the extracellular fluid. This might explain the results recorded here concerning the decrease in tissue EC-SOD activity and increase in plasma of breast cancer patients.

• YAKHAK HOEJI
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• v.43 no.5
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• pp.598-605
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• 1999

The hot water and mathanol extracts of Artemisia argyi showed considerable cytotoxicities against H9(ATCC HTB 176) cancer cell with IC50 values of $48.6{\;}\mu\textrm{g}/ml$ and $51.9{\;}\mu\textrm{g}/ml$, respectively. These cytotoxicities were found to be dependent on the extract concentrations and culture days. CuZnSOD and MnSOD activities were significantly increased in the cytoplasm and mitochondria fractions of cancer cell, and media in the presence of Artemisia argyi. Such enhanced SOD activities were generally in the range of two to threefolds. In contrast to SOD, catalase and glutathione peroxidase activities were not detected at all. These results suggest that Artemisia argyi have generated $O_2^-$ in the mitochondria and cytoplasm of H9 cancer cell with concurrent induction of CuZnSOD and MnSOD in situ, which dismutate $O_2^-{\}to{\;}H_2O_2$. Without coordinated actions of catalase and/or glutathione peroxidase $H_2O_2$ is easily converted to very toxic OH and these reactive oxygen species together might have induced necrosis and/or apoptosis of H9 cell.

• FLOWER RESEARCH JOURNAL
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• v.19 no.4
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• pp.269-273
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• 2011

This study was conducted to investigate the resistance to abiotic stress in the progeny obtained by a cross between NDPK2-transgenic line (NDPK2-7-1) and MnSOD (SOD2) transgenic line (SOD2-2-1-1-35) to develop transgenic petunia highly resistant to environmental stress. At the treatment of 100 and $200{\mu}M$ methyl viologene (MV), the progeny was significantly less damaged than its parental plants (SOD2- or NDPK2-transgenic lines) as well as non-transgenic plants, implying its resistance to oxidative stress enhanced than SOD2- or NDPK2-transgenic plants. In an expression of 11 quantitative traits, the progeny remained similar to control plants, although it infrequently displayed slightly longer or wider than non-transgenic control plants. In the color and shape of flowers, there was no significant difference between the progeny and its parents or non-transgenic control.

• Bulletin of the Korean Chemical Society
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• v.32 no.7
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• pp.2405-2409
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• 2011

The psychrophilic bacteria Psychromonas arctica survives at subzero temperatures by having adapted several protective mechanisms against freezing and oxidative stresses. Many reactive oxygen species are likely generated in P. arctica as a result of reduced metabolic turnover rates. A previous study identified the pasod gene for superoxide dismutase from P. arctica using a series of PCR amplifications. Here, upon cloning into a His-tag fused plasmid, the sod gene from P. arctica (pasod) was successfully expressed by IPTG induction. His-tagged PaSOD was subsequently purified by $Ni^{2+}$-NTA affinity chromatography. The purified PaSOD exhibited a higher SOD activity than that of Escherichia coli (EcSOD) at all temperatures. The difference in activity between PaSOD and EcSOD becomes even more significant at 4$^{\circ}C$, indicating that PaSOD plays a functional role in the cold adaptation of P. arctica in the Arctic.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.229-234
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• 2006

We have generated transgenic mice that expressed mouse extracellular superoxide dismutase (EC-SOD) in their skin. In particular, the expression plasmid DNA containing human keratin K14 promoter was used to direct the keratinocyte-specific transcription of the transgene. To compare intron-dependent and intron-independent gene expression, we constructed two vectors. The vector B, which contains the rabbit -globin intron 2, was not effective for mouse EC-SOD overexpression. The EC-SOD transcript was detected in the skin, as determined by Northern blot analysis. Furthermore, EC-SOD protein was detected in the skin tissue, as demonstrated by Western blot analysis. To evaluate the expression levels of EC-SOD in various tissues, we purified EC-SOD from the skin, lungs, brain, kidneys, livers, and spleen of transgenic mice and measured its activities. EC-SOD activities in the transgenic mice skin were approximately 7 fold higher than in wild-type mice. These results suggest that the mouse overexpressing vector not only induces keratinocyte-specific expression of EC-SOD, but also expresses successfully functional EC-SOD. Thus, these transgenic mice appeared to be useful for the expression of the EC-SOD gene and subsequent analysis of various skin changes, such as erythema, inflamation, photoaging, and skin tumors.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.201-206
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• 1998

To develop the fruits of cucumber (Cucumis sativus L.) producing high yields of superoxide dismutase (SOD), the MnSOD cDNA from pea (Pisum sativum) under the control of the cauliflower mosaic virus 35S promoter was introduced into cucumber using Agrobacterium tumefaciens (strain LBA 4404)-mediated transformation. The kanamycin-resistant shoots were selected on the selection medium containing MS basal salt, 1.0 mg/L zeatin, 0.1 mg/L IAA, 300 mg/L claforan, and 100 mg/L kanamycin. After 6 weeks of culture on the selection medium, the shoots were transferred to MS medium containing 0.2 mg/L NAA to induce roots. PCR analysis using the primers for neomycin phosphotransferase (NPTII) gene revealed that three plantlets were transformed. The fruits of one transgenic plant had approximately 3.2-fold higher SOD activity than those of non-transgenic plants. MnSOD isoenzyme band was strongly detected on native gel in fruits of transgenic plants.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1484-1487
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• 1998

Superoxide dismutase (SOD)-like activities of sixteen kinds of fruits, vegetable juice and commercial concentrates were measured by pyrogallol autoxidation method. The changes in SOD-like activity by heat treatment and the increase in SOD-like activity of apple juice mixed with fruits and vegetables were investigated. SOD-like activity of broccoli juice was 41.7%, the highest value among tested sample. SOD-like activities of strawberry juice, carrot concentrate, kiwi juice, radish juice and apple juice were 30.2, 30.0, 27.6, 26.7, 24.1 and 14.6%, respectively. SOD-like activity was increased generally after heat treatment at $95^{\circ}C$ until 20 min. SOD-like activity of apple juice was increased $20{\sim}35%$ by mixing with 20% of carrot concentrate, kiwi juice, strawberry juice, broccoli juice, respectively and particularly was increased 48% by mixing with 20% of raddish juice.

• BMB Reports
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• v.32 no.6
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• pp.585-593
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• 1999

The water and methanol extracts of Artemisia argyi showed significant cytotoxicities against J774A.1 cells but not so much against normal leukocytes. The cytotoxicities were found to be dependent on the extract concentration and the incubation time. The concentration of water and methanol extracts inhibiting 50% of cell proliferation ($IC_{50}$) were estimated to be 44.2 mg/ml and 71.6 mg/ml, respectively. In the presence of Artemisia argyi water extract, total superoxide dismutase (CuZnSOD and MnSOD) activities of media, cytoplasmic and mitochondrial fractions of J774A.1 cells increased in accordance with cytotoxicity. MnSOD was found to be the main component of enhanced total SOD activities, particulary in the mitochondrial fraction. In contrast to SOD, catalase and glutathione peroxidase (GPx) were not found in any instance of the current investigation. In addition, substantial amount of $O_2^-$ appeared to be generated in the mitochondrial fraction under the influence of Artemisia argyi. All data put together, it is postulated that Artemisia argyi extracts seem to stimulate $O_2^-$ generation in mitochondria of J774A.1 cells with concomitant increases of SODs. Since $H_2O_2$, the reaction product of SOD on $O_2^-$, is known to be readily converted to very toxic $OH{\cdot}$ in the absence of catalase and/or GPx cooperation, toxicity derived from ROS such as $O_2^-$, $H_2O_2$, and $OH{\cdot}$ may be the main cause of necrosis and/or apoptosis of J774A.1 cells.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.215-221
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• 1994

Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1881-1890
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• 2016

Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.