디젤분진이 체세포에서의 DNA 손상에 미치는 영향

허찬,김남이,정규혁,문창규,허문영 한국환경독성학회 2004 환경독성보건학회지 Vol.19 No.4

Diesel exhaust panicle (<25μm, DEP_(2.5)) is known to be probarbly carcinogenie (IARC group 2A) DEP_(2.5) contains organic compounds such as polycyclicatomatic hydrocarbon (PAH), heterocyclic compounds, phenols, and nitroarenes Reactive oxygen species (ROS) are generated by DEP_(2.5) without any biological activation system Therefore, an alternative mechanism by which DEP_(2.5) could be carcinogenic is known by the generation of oxidative DNA damage The aim of this study was to investigate genotoxic effects of DEP_(2.5) using single cell gel electrophoresis In order to evaluate the mechanisms of DEP_(2.5) genotoxicity, the rat microsome mediated and DNA repan enzyme treated comet assays together with routine comet assay were performed DEP_(2.5) was collected from diesel engine bus and dichloiomethane extract was obtained The organic extract of DEP_(2.5) revealed DNA damage itself in NIH/3T3 cells And it showed both oxidative and microsome mediated DNA damages Vitamin C as an model antioxidant reduced DNA damage in endonuclase Ⅲ treated comet assay One of flavonoid, galangin as a CYP[A] inhibitor reduced DNA damage in the presence of S-9 mixture Our results show that DEP_(2.5) ate genotoxic and a great source of oxidative stress, but antioxidants can significantly reduce oxidative DNA damages And DEP_(2.5) may contain indirect mutagens which can be inhibited by CYP inhibitors.

Identification of a Tumor-Associated Autoantibody against Aminoimidazole Carboxamide Ribonucleotide Transformylase/Inosine Monophosphate Cyclohydrolase (ATIC) in Hepatocellular Carcinoma (HCC) Mouse Model and Its Application to Diagnosis of HCC

Chang-Kyu Heo,Mi-Kyung Woo,Hai-Min Hwang,Ah Ruem Kim,Dae-Yeul Yu,Ju Yeon Lee,Jong Shin Yoo,Chulhun Chang,Jeong Heon Ko,Eun-Wie Cho 한국당과학회 2012 한국당과학회 학술대회 Vol.2012 No.1

Autoantibodies, which are generated by immune system recognizing the presence of the abnormal tumor-associated antigens, are promising biomarkers for the early detection of tumors. Recently, we established a B cell hybridoma pool derived from HBx transgenic mouse, as a source of tumor-associated autoantibodies without using any extracellular antigens, and have characterized the specific target antigens against them. XC154 autoantibody, one of them, has been investigated in this study and its target antigen was identified by mass spectrometric analysis as aminoimidazole carboxamide ribonucleotide transformylase / inosine monophosphate cyclohydrolase (ATIC) that catalyzes the last two steps of de novo purine biosynthesis. A specific mimotope against XC154 autoantibody was screened from the cyclic random hepta-peptide phage library and, using mimotope display M13 phage as a coating antigen for ELISA, we could distinguish patients with hepatocellular carcinoma (HCC) vs. normal subjects with 86.96% sensitivity and 88.24% specificity. These results imply that anti-ATIC autoantibody is induced in patients with HCC and detection of anti-ATIC autoantibody can be used for the diagnosis of HCC. Now, several attempts have been performed to establish a more reliable and convenient assay to detect tumor-assocoated autoantibodies using recombinant proteins displaying cycling peptide instead of mimotope display M13 phage and the usefulness of improved assay will be discussed.

Identification of autoantibody against cytokeratin 8/18 complex in H-ras12V tumor model mouse and its application to diagnosis of human breast cancer

Chang-Kyu Heo,Hae-Min Hwang,Dae-Yeul Yu,Ju Yeon Lee,Jong-ShinYoo,Hyang Sook Yoo,Jeong Heon Ko,Jin-Man Kim,Sejeong Oh,Eun-Wie Cho 한국당과학회 2011 한국당과학회 학술대회 Vol.2011 No.1

Autoantibodies, which are generated by immune system recognizing the presence of the abnormal tumor-associated antigens, are promising biomarkers for the early detection of tumors. Recently, we established hundreds of B cell hybridomas from H-ras12V transgenic mouse, a typical tumor model system, as a source of tumor-associated autoantibodies without using any extracellular antigens and have characterized specific target antigens against them. TAB-K94 monoclonal antibody, one of tumor-associated autoantibodies obtained from H-ras12V transgenic mouse, was investigated in this study and its target antigen was identified as cytokeratin 8/18 (CK8/18) complex, an intermediate filament (IF) protein complex of epithelial cells which is involved in cell motility and cancer progression. To establish a method for the detection of autoantibodies against CK8/18 complex in tumor patients sera, a specific mimotope against TAB-K94 autoantibody was screened from the cyclic random hepta-peptide phage library and, by enzyme-linked immunosorbent assay (ELISA) using it as a binder of anti-CK8/18 autoantibodies, we could distinguish breast cancer patients vs. normal subjects with 50.00 % sensitivity and 82.61 % specificity. These results imply that anti-CK8/18 autoantibody is induced in a certain group of breast cancer patients and detection of anti-CK8/18 autoantibody would be useful for the diagnosis of breast cancer.

Identification of a mimotope for circulating anti-cytokeratin 8/18 antibody and its usage for the diagnosis of breast cancer

HEO, CHANG-KYU,HWANG, HAI-MIN,RUEM, AH,YU, DAE-YEUL,LEE, JU YEON,YOO, JONG SHIN,KIM, IN GYU,YOO, HYANG SOOK,OH, SEJEONG,KO, JEONG HEON,CHO, EUN-WIE D.A. Spandidos 2013 International journal of oncology Vol.42 No.1

<P>A novel circulating tumor-associated autoantibody, K94, obtained from a hepatocellular carcinoma (HCC) mouse model was characterized. The target antigen of K94 autoanti-body was expressed in various tumor cell lines including liver cancer, and its secretion was detectable using MCF-7 breast carcinoma cells. Proteomic analysis revealed that the protein bands reactive to K94 included cytokeratin (CK) 8 and 18, which are known to be related to tumorigenesis and form a heterotypic complex with each other. However, K94 showed no activity toward CK8 or CK18 separately. The epitope of the K94 antibody was only presented by a complex between CK8 and CK18, which was confirmed by analysis using recombinant CK8 and CK18 proteins. To formulate an assay for anti-CK8/18 complex autoantibody, a mimotope peptide reactive to K94 was selected from loop-constrained heptapeptide (-CX<SUB>7</SUB>C-) display phage library, of which sequence was CISPDAHSC (K94p1). A mimotope enzyme-linked immunosorbent assay (ELISA) using phage-displayed K94p1 peptide as a coating antigen was able to discriminate breast cancer (n=30) patients from normal subjects (n=30) with a sensitivity of 50% and a specificity of 82.61%. CA15.3 was detected at very low levels in the same breast cancer subjects and did not discriminate breast cancer patients from normal subjects, although it is a conventional biomarker of breast cancer. These results suggest that a mimotope ELISA composed of K94p1 peptide may be useful for the diagnosis of breast cancer.</P>

Invited Mini Review : Tumor-associated autoantibodies as diagnostic and prognostic biomarkers

( Chang Kyu Heo ),( Young Yil Bahk ),( Eun Wie Cho ) 생화학분자생물학회(구 한국생화학분자생물학회) 2012 BMB Reports Vol.45 No.12

In the process of tumorigenesis, normal cells are remodeled to cancer cells and protein expression patterns are changed to those of tumor cells. A newly formed tumor microenvironment elicits the immune system and, as a result, a humoral immune response takes place. Although the tumor antigens are undetectable in sera at the early stage of tumorigenesis, the nature of an antibody amplification response to antigens makes tumor-associated autoantibodies as promising early biomarkers in cancer diagnosis. Moreover, the recent development of proteomic techniques that make neo-epitopes of tumor-associated autoantigens discovered concomitantly has opened a new area of ``immuno-proteomics``, which presents tumor-associated autoantibody signatures and confers information to redefine the process of tumorigenesis. In this article, the strategies recently used to identify and validate serum autoantibodies are outlined and tumor-associated antigens suggested until now as diagnostic/prognostic biomarkers in various tumor types are reviewed. Also, the meaning of autoantibody signatures and their clinical utility in personalized medicine are discussed. [BMB Reports 2012; 45(12): 677-685]

Enhanced Anti-inflammatory Effects of γ-irradiated Pig Placenta Extracts

Kang Chang Kim,Jae Hyeok Heo,Jong Kwang Yoon,Yuyeon Jang,Youn Kyu Kim,Chang Kyu Kim,Yu Kyung Oh,Young Bong Kim 한국축산식품학회 2015 한국축산식품학회지 Vol.35 No.3

Porcine placenta extract (PPE) is known to possess anti-inflammatory properties owing to its high concentration of bioactive substances. However, the need to eliminate blood-borne infectious agents while maintaining biological efficacy raises concerns about the optimal method for sterilizing PPE. Therefore, the objective of this study was to compare the effects of the standard pressurized heat (autoclaving) method of sterilization with γ-irradiation on the anti-inflammatory effects of PPE. The anti inflammatory actions of these two preparations of PPE were evaluated by measuring their inhibitory effects on the production of NO, the expression of iNOS protein, and the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA in lipopolysaccharide-stimulated RAW 264.7 cells. Compared with autoclaved PPE, γ-irradiated PPE showed significantly greater inhibition of NO production and iNOS protein expression, and produced a greater reduction in the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA. These results provide evidence that the sterilization process is crucial in determining the biological activity of PPE, especially its anti-inflammatory activity. Collectively, our data suggest that γ-irradiated PPE acts at the transcriptional level to effectively and potently suppresses the production of NO and the expression of pro-inflammatory cytokines.

Enhanced Anti-inflammatory Effects of γ-irradiated Pig Placenta Extracts

Kim, Kang Chang,Heo, Jae Hyeok,Yoon, Jong Kwang,Jang, Yuyeon,Kim, Youn Kyu,Kim, Chang-Kyu,Oh, Yu-Kyung,Kim, Young Bong Korean Society for Food Science of Animal Resource 2015 한국축산식품학회지 Vol.35 No.3

Porcine placenta extract (PPE) is known to possess anti-inflammatory properties owing to its high concentration of bioactive substances. However, the need to eliminate blood-borne infectious agents while maintaining biological efficacy raises concerns about the optimal method for sterilizing PPE. Therefore, the objective of this study was to compare the effects of the standard pressurized heat (autoclaving) method of sterilization with γ-irradiation on the anti-inflammatory effects of PPE. The anti-inflammatory actions of these two preparations of PPE were evaluated by measuring their inhibitory effects on the production of NO, the expression of iNOS protein, and the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA in lipopolysaccharide-stimulated RAW 264.7 cells. Compared with autoclaved PPE, γ-irradiated PPE showed significantly greater inhibition of NO production and iNOS protein expression, and produced a greater reduction in the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA. These results provide evidence that the sterilization process is crucial in determining the biological activity of PPE, especially its anti-inflammatory activity. Collectively, our data suggest that γ-irradiated PPE acts at the transcriptional level to effectively and potently suppresses the production of NO and the expression of pro-inflammatory cytokines.

Noninvasive Monitoring of Bleomycin-induced Lung Injury in Rats Using Pulmonary Function Test

Yang, Mi-Jin,Yang, Young-Su,Kim, Yong-Bum,Cho, Kyu-Hyuk,Heo, Jeong-Doo,Lee, Kyu-Hong,Song, Chang-Woo Korean Society of ToxicologyKorea Environmental Mu 2008 Toxicological Research Vol.25 No.2

The single intratracheal instillation (ITI) of bleomycin (BLM) is a widely used method for inducing experimental pulmonary fibrosis in rat model. In the present study, pulmonary function tests (PFTs) of tidal volume ($V_T$), minute volume ($V_M$), and respiratory frequency ($F_R$) have been applied to study their possibility as a tool to monitor the progress of BLM-induced lung injury in rat model. Rats were treated with a single ITI of BLM (2.5 mg/kg) or saline (control). Animals were euthanized at 3, 7, 14, 21, and 28 days post-ITI. Lung toxicity effects were evaluated by inflammatory cell count, lactate dehydrogenase (LDH) activity in the bronchoalveolar lavage fluid (BALF), and light microscopic examination of lung injury. The PFT parameters were measured immediately before the animals were sacrificed. BLM treatment induced significant cellular changes in BALF-increase in number of total cells, neutrophils, and lymphocytes along with sustained increase in number of macrophages compared to the controls at days 3, 7, and 14. BALF LDH level was significantly increased compared to that in the controls up to day 14. On day 3, infiltration of neutrophils was observed in the alveolar spaces. These changes developed into marked peribronchiolar and interstitial infiltration by inflammatory cells, and extensive thickening of the interalveolar septa on day 7. At 14, 21, and 28 days, mild peribronchiolar fibrosis was observed along with inflammatory cell infiltration. The results of PFT show significant consistencies compared to the results of other toxicity tests. These data demonstrate that the most suitable time point for assessing lung fibrosis in this model is 14 days post-ITI of BLM based on the observation of fibrosis at 14, 21, and 28 days. Further, the progress of lung injury can be traced by monitoring the PFT parameters of $F_R$, $V_T$, and $V_M$.

Inhibition of translocation of p65 by extracts of Campsis grandiflora in ROS 17/2.8 cells activated with cytokines

Chang Ki Churl,Oh Hwa Min,Kang Young Jin,Park Min Kyu,Lee Young Soo,Heo Ja Myung,Lee Jae Heun,Seo Han Geuk,Yun-Choi Hye Sook 대한약학회 2004 大韓藥學會 總會 및 學術大會 Vol.2004 No.1

Enhanced bone regeneration with a gold nanoparticle-hydrogel complex

Heo, Dong Nyoung,Ko, Wan-Kyu,Bae, Min Soo,Lee, Jung Bok,Lee, Deok-Won,Byun, Wook,Lee, Chang Hoon,Kim, Eun-Cheol,Jung, Bock-Young,Kwon, Il Keun The Royal Society of Chemistry 2014 Journal of Materials Chemistry B Vol.2 No.11

<P>Gold nanoparticles (GNPs) are widely used in diagnostics, drug delivery, biomedical imaging, and photo-thermal therapy due to their surface plasmon resonance, fluorescence, and easy-surface functionalization. According to recent studies, GNPs display a positive effect on the osteogenic differentiation of mesenchymal stem cells (MSCs) and MC3T3-E1 osteoblast-like cells. The aim of this study was to develop a new approach for bone tissue regeneration based on the utilization of a biodegradable hydrogel loaded with GNPs. We have used photo-curable gelatin hydrogels (Gel) in order to provide a proof of principle of GNPs in regeneration strategies for bone tissue repair. We have investigated the effects of these Gel-GNP composite hydrogels both <I>in vitro</I> and <I>in vivo</I>. The <I>in vitro</I> results showed that the hydrogels loaded with GNPs promote proliferation, differentiation, and alkaline phosphate (ALP) activities of human adipose-derived stem cells (ADSCs) as they differentiate towards osteoblast cells in a dose-dependent manner. Moreover, the <I>in vivo</I> results showed that these hydrogels loaded with high concentrations of GNPs had a significant influence on new bone formation. Through these <I>in vitro</I> and <I>vivo</I> tests, we found that the Gel-GNP can be a useful material for bone tissue engineering.</P>