• 한국환경성돌연변이발암원학회지
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• 제9권1호
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• pp.13-22
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• 1989

The effect of novobiocin (NOV), and inhibitor of topoisomerase II, on ethyl methanesulfonate (EMS)-or bleomycin (BLM)-induced DNA repair synthesis was examined during the cell cycle of Chinese hamster ovary (CHO)-K1 cells. Three assays were employed in this study: cell survival, alkaline elution and unscheduled DNA synthesis. EMS was effective at killing CHO cells in G1 phase, wheras BLM preferentially killed cells in G2 and S phases. EMS induced the much more amount of DNA damage in G1 phase, while BLM induced in G2 phase than the other phases. The both of pre- and post-treatment with BOV inhibitied EMS- or BLM-induced DNA repair synthesis in G1 and G2 phases, and pretreatment with NOV inhibited more effectively than the post-treated group. These results suggested that CHO cells exhibited a differential sensitivity to cell lethality and DNA damage in relation to cell cycle according to used chemical agents, and that DNA topoisomerase II participated in an initial stage of DNA repair.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.74-74
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• 2018

Abeliophyllum distichum is Korea Endemic Plants and its genetic resources found from Korea only. Bioactivities of A. distichum such as antioxidant, anti-cancer, and anti-inflammatory studies have been proved through many researches. Whereas, there are no studies on the biological activity of its callus extracts. In this study, we investigated the antioxidant activities of callus extracts derived from A. distichum and its inhibitory effect on oxidative DNA damage. The antioxidant activities were assessed using radical scavenging assays with DPPH, ABTS, and reducing power assay and the inhibitory effects on oxidative DNA damage were measured using ${\varphi}-174$ RF I plasmid DNA cleavage assay. In addition, callus extracts derived from A. distichum showed high antioxidant acitivties and no cytotoxicity in NIH/3T3. Also, it has significantly suppressed expression of ${\gamma}$-H2AX and p53 protein and mRNA levels in NIH/3T3 cells exposed to oxidative stress. Therefore, the callus extracts derived from A. distichum has potential antioxidant activity that can provide protective effects against the oxidative DNA damage caused by free radicals. This study suggest that it is valuable as cosmetics and medicine for antioxidant and cancer preventive materials.

• 대한본초학회지
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• 제30권2호
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• pp.31-36
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• 2015

Objectives : In this study, we demonstrated the antioxidant activities and the inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Sophorae Flos and Sophorae fructus. Methods : Sophorae Flos and Sophorae fructus were extracted with methanol(MeOH) and divided to Petroleum ether, Ethyl acetate(EtOAC) and Water fraction. The antioxidant activities were conducted by the 1,1-diphenyl-2-picryl hydrazyl(DPPH) radical, 2, 2'-Azine-bis(3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt(ABTS) radical scavenging assay, $Fe^{2+}$ chelating assay and Reducing power assay. The inhibitory effect of DNA damage were characterized on ${\varphi}$ X-174 RF I plasmid DNA cleavage assay. In addition, we analyzed the Total phenol contents and the Vitamin C contents of Sophorae Flos and Sophorae fructus. Results : The results of DPPH were 92.71% and 94.72%, ABTS were 87.16% and 62.44%, and $Fe^{2+}$ chelating were 95.81% and 85.11% at $200{\mu}g/m{\ell}$ of Sophorae Flos and Sophorae fructus respectively. The Sophorae Flos showed stronger effect than Sophorae fructus in Reducing Power assay. Total phenol content was 111.77 mg/g and 122.54 mg/g, and Vitamin C content was 2.59 mg/g and 3.03 mg/g. Also both Sophorae Flos and Sophorae fructus have inhibitory antioxidant effect on ${\varphi}$ X-174 RF I plasmid DNA cleavage assay. Conclusions : Over all, this study suggests that Sophorae Flos and Sophorae fructus can be used as not only effective antioxidant but also natural medicine.

• 한국해양바이오학회지
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• 제14권1호
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• pp.33-42
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• 2022

Tacrolimus, a type of macrolide produced by Streptomyces tsukubaensis, is widely used as an immunosuppressant. However, continuous exposure to tacrolimus causes oxidative stress in normal cells, ultimately inducing cell injury. Therefore, this study investigated whether luthione, a reduced glutathione, could inhibit tacrolimus-induced cytotoxicity in olive flounder (hirame) natural embryo (HINAE) cells. According to the results, luthione significantly inhibited tacrolimus-induced reduction in cell viability in a concentration-dependent manner. Additinally, although luthione unaffected autophagy by tacrolimus, tacrolimus-induced apoptosis was significantly suppressed in the presence of luthione. Luthione also markedly blocked DNA damage in tacrolimus-treated HINAE cells, associated with the inhibition of reactive oxygen species (ROS) generation. Additionally, tacrolimus cytotoxicity in HINAE cells was correlated with increased inflammatory response, also attenuated by luthione. Collectively, these results show that at least luthione protects HINAE cells against tacrolimus-induced DNA damage, apoptosis, and inflammation, but not autophagy, by scavenging ROS. Although additional in-vivo studies are required, this study's results can be used as a basis for utilizing luthione to reduce the toxicity of fish cells caused by excessive immune responses.

• 한국식품위생안전성학회지
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• 제17권2호
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• pp.106-116
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• 2002

양강추출물 (Alpinia officinarum, 70% ethanol extract)과 함유 주성분인 galangirn은 프리라디칼소거작용과 지질과산화억제활성을 나타내었으며, $H_2O$$_2$ 또는 KO$_2$유도 세포독성에 대해서도 억제적으로 작용하여 세포보호효과를 나타내었다. DNA single strand breakage와 같은 산화적 스트레스에 대해서도 보호작용을 하고 있다. 또한, 마우스 소핵시험에 의하여 adriamycin과 같은 superoxide 우발물질에 의한 염색체 수준에서의 손상에 대해서도 억제효과를 나타내었다. 양강추출물은 galangin을 유효성분으로하여 산소프리라디칼들에 의한 산화적 손상에 억제적으로 작용하는 기전으로 산화적 스트레스에 대한 항노화, 암예방제로서의 응용가능성이 높은 추출물로서 향후 기능성 식품으로서의 응용가능성이 높을 것으로 판단되었다.

• 약학회지
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• 제49권3호
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• pp.249-254
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• 2005

The ethanol extracts of the mixed vegetables (Bioactive Vegetables, BV) and the mixed fruits (Bioactive Fruits, BF) were evaluated for their in vivo antioxidant activities. Four weeks treatment of oral administration was performed to mice. A $KBrO_3$ as a potent oxidant was used to induce the oxidative stress for in vivo experiment. BV and BF were shown to possess the significant inhibitory effect of lipid peroxidation as measured by the level of malondialdehyde (MDA) formation although the potencies were not higher than those of well-known antioxidants such as vitamin C, trolox and quercetin. Furthermore, BV and BF inhibited DNA damage assessed by single cell gel electrophoresis (comet assay) and reduced the micronucleated reticulocyte (MNRET) formation of peripheral blood. Antioxidants tested also revealed potent inhibitory activities higher than BV and BF. These antigenotoxic activity profiles were similar to that of abovementioned inhibition of lipid peroxidation. Therefore, BV and BF having mild antioxidant activity as functional food candidates may be useful natural antioxidants by the inhibiting of lipid peroxidation and the protecting oxidative DNA and chromosomal damage.

• 약학회지
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• 제48권2호
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• pp.111-116
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• 2004

The ethanol extracts of mixed vegetables (Bioactive Vegetables, BV), mixed fruits (Bioactive Fruits, BF) and their liquid formulation (Chungpae Plus , CP) were evaluated for their antioxidative and antigenotoxic activities. They were shown to possess the significant free radical scavenging effect against 1,1-diphenyl-2-picryl hydrazine (DPPH) radical generation and were revealed to show the inhibitory effect of lipid peroxidation as measured by malondialdehyde (MDA) formation although the potencies were not higher than those of vitamin C. They did not possess any pro-oxidant effect on bleomycin-Fe(III) dependent DNA degradation, whereas vitamin C showed strong pro-oxidant effect. Furthermore, oral administration of BV and BF inhibited micronucleated reticulocyte (MNRET) formation of mouse peripheral blood induced by KBrO3 treatment in vivo. CP also showed significant inhibition under same experimental condition. Therefore, the liquid formulation (CP) containing BV and BF may be a useful natural antioxidative and antigenotoxic agent by scavenging free radicals, inhibition of lipid peroxidation and protecting chromosomal damage.

• 한국임상수의학회지
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• 제21권3호
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• pp.253-258
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• 2004

DNA double-strand break (DSB) is a serious treat for the cells including mutations, chromosome rearrangements, and even cell death if not repaired or misrepaired. Ku heterodimer regulatory DNA binding subunits (Ku70/Ku80) bound to double strand DNA breaks are able to interact with 470-kDa DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and the interaction is essential for DNA-dependent protein kinase (DNA-PK) activity. The Ku80 mutants were designed to bind Ku70 but not DNA end binding activity and the peptides were treated in breast cancer cells for co-therapy strategy to see whether the targeted inhibition of DNA-dependent protein kinase (DNA-PK) activity sensitized breast cancer cells to ionizing irradiation or chemotherapy drug to develop a treatment of breast tumors by targeting proteins involved in damage-signaling pathway and/or DNA repair. We designed domains of Ku80 mutants, 26 residues of amino acids (HN-26) as a control peptide or 38 (HNI-38) residues of amino acids which contain domains of the membrane-translocation hydrophobic signal sequence and the nuclear localization sequence, but HNI-38 has additional twelve residues of peptide inhibitor region. We observed that the synthesized peptide (HNI-38) prevented DNA-PKcs from binding to Ku70/Ku80, resulting in inactivation of DNA-PK complex activity in breast cancer cells (MDA-465 and MDA-468). Consequently, the peptide treated cells exhibited poor to no DNA repair, and became highly sensitive to irradiation or chemotherapy drugs. The growth of breast cancer cells was also inhibited. These results demonstrate the possibility of synthetic peptide to apply breast cancer therapy to induce apoptosis of cancer cells.

• Journal of Preventive Medicine and Public Health
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• 제30권3호
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• pp.555-566
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• 1997

This study was carried out to evaluate the cytotoxicity of rock wool fibers(RWFs) such as cell division disturbance, chromosomal and DNA damage, and mutagenicity using cultured cells. RWFs were the man made mineral fibers. In order to find the correlation between the cytotoxicity of RWFs and the phagocytic capacity of cells, the phagocytic processes were observed using scanning electron microscope. Cell division disturbance by RWFs was evaluated by the formation of multinucleated giant cells. The chromosomal damage was evaluated by the micronucleus formation. For the evaluation of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8-OH-dG) formation was measured utilizing calf thymus DNA. Mutagenicity was determined by the point mutation of HGPRT and the effect of RWFs on cell transformation was also observed. 1. Compared with the results of chrysotile, RWFs were no or little effect on the cell growth according to the results done by the tests of cell proliferation inhibition and relative plating efficiency. 2. The frequency of multinucleated giant cell formation was increased by the treatment of RWFs and it was dose-dependent. However, the effect of RWFs was weaker than that of chrysotile. 3. The number of micronuclei formed in the RWFs treated cells was between those of cells treated with chrysotile and those of untreated cells. 4. The 2 fold increase in the formation of 8-OH-dG in calf thymus DNA was observed in the cells treated with RWFs in the presence of $H_2O_2$. On the other hand, chrysotile had no effect on the 8-OH-dG formation. 5. RWFs had no effect on the HGPRT point mutation and cell transformation. These results showed that RWFs could induce chromosomal damage, cell division disturbance and oxidative DNA damage in the RWFs treated cells.

• The Korean Journal of Physiology and Pharmacology
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• 제7권1호
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• pp.9-14
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• 2003

Recent studies indicated that cancer cells become resistant to ionizing radiation (IR) and chemotherapy drugs by enhanced DNA repair of the lesions. Therefore, it is expected to increase the killing of cancer cells and reduce drug resistance by inhibiting DNA repair pathways that tumor cells rely on to escape chemotherapy. There are a number of key human DNA repair pathways which depend on multimeric polypeptide activities. For example, Ku heterodimer regulatory DNA binding subunits (Ku70/Ku80) on binding to double strand DNA breaks (DSBs) are able to interact with 470-kDa DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and are essential for DNA-dependent protein kinase (DNA-PK) activity. It has been known that DNA-PK is an important factor for DNA repair and also is a sensor-transmitting damage signal to downstream targets, leading to cell cycles arrest. Our ultimate goal is to develop a treatment of breast tumors by targeting proteins involved in damage-signaling pathway and/or DNA repair. This would greatly facilitate tumor cell cytotoxic activity and programmed cell death through DNA damaging drug treatment. Therefore, we designed a domain of Ku80 mutants that binds to Ku70 but not DNA end binding activity and used the peptide in co-therapy strategy to see whether the targeted inhibition of DNA-PK activity sensitized breast cancer cells to irradiation or chemotherapy drug. We observed that the synthesized peptide (HNI-38) prevented DNA-PKcs from binding to Ku70/Ku80, thus resulting in inactivation of DNA-PK activity. Consequently, the peptide treated cells exhibited poor to no DNA repair, and became highly sensitive to IR or chemotherapy drugs, and the growth of breast cancer cells was inhibited. Additionally, the results obtained in the present study also support the physiological role of resistance of cancer cells to IR or chemotherapy.