• 디지털융복합연구
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• 제17권1호
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• pp.443-447
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• 2019

방향성탄화수소 수용체(Arylhydrocarbon Receptor, AhR)은 리간드에 의해 활성화되어 체내 외래물질의 대사를 조절하는 전사인자다. 생체 내에서 AhR의 생리학적 역할은 오랜 기간 연구되어 왔으나 활성화를 유발하지 않는 길항제를 비롯하여 유효한 화학적 도구가 아직 개발되지 않아 기능 연구가 제한적이다. AhR이 다양한 질병의 발병기전에서 중요한 역할을 수행한다는 것이 보고됨에 따라 약물 표적으로서 유효하다고 판단되나 치료나 예방을 위한 유효한 약물은 아직 개발되지 않았다. 길항제로 알려진 화합물들은 낮은 농도에서는 활성이 있어 연구 목적으로 활용되고 있으나 높은 농도에서는 방향성 탄화수소를 활성화하는 부분적 agonist로 작용한다. 이에 AhR 활성화를 유도하지 않는 순수한 길항제의 개발이 필요하다. 본 연구에서는 이미 알려진 AhR 길항제의 골격인 phenyldiazenylphenylpicolinamide의 세 고리구조 중 두 고리구조에 존재하는 메틸 기들을 변형하여 활성을 평가하는 구조-활성 관계 연구를 통하여 새로운 길항제를 개발하고자 하였다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.187-187
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• 2001

Since hypoxia-inducible factor-lalpha (HIF-lalpha) and the arylhydrocarbon receptor (AhR) shared the AhR nuclear translocator (Arnt) for hypoxia- and AhR-mediated signaling, respectively, it was possible to establish the hypothesis that hypoxia could regulate Cyp1a1 expression.(omitted)

• 한국융합학회논문지
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• 제10권1호
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• pp.285-290
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• 2019

방향성 탄화수소 수용체(Aryl Hydrocarbon Receptor, AHR)은 리간드에 의해 활성화되어 체내 외래물질의 대사를 조절하는 전사인자다. 생체 내에서 AHR의 생리학적 역할은 오랜 기간 연구되어 왔으나 길항제를 비롯한 적절한 화학적 도구의 부재로 그 역할 규명이 제한되어 있다. AHR이 다양한 질병의 발병기전에 관여되어 있다는 것이 밝혀짐에 따라 유효한 약물 표적으로 인식되고 있으나 치료나 예방을 위한 유효한 약물은 아직 개발되지 않았다. 길항제로 알려진 화합물들은 낮은 농도에서는 길항활성이 있어 연구 목적으로는 활용되고 있으나 높은 농도에서는 AHR을 활성화하는 부분적 agonist로 작용한다. 이에 AHR 활성화를 유도하지 않는 순수한 길항제의 개발이 필요하다. 본 연구에서는 이미 알려진 AHR 길항제인 Resveratrol과 CH223191의 구조를 기반으로 phenyldiazenylanline 구조를 설계하였고 이를 골격으로 다양한 유도체를 합성하고 화학적 구조와 생물학적 활성의 상관 관계에 대한 융합 연구를 통하여 신규 AHR 길항제를 도출하였다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.114-114
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• 2001

Since hypoxia-inducible factor-lalpha (HIF-lalpha) and the arylhydrocarbon receptor (AhR) shared the AhR nuclear translocator (Arnt) for hypoxia- and AhR-mediated signaling, respectively, it was possible to establish the hypothesis that hypoxia could regulate Cyplal expression. In order to understand the mechanism of Cyplal gene expression, we demonstrated here that hypoxic agents such as cobalt chloride, desferrioxarnine, and picolinic acid reduced the TCDD induced Cyplal promoter activity based on the determination of luciferase activity in Hepa I cells transfected with pmCypla1-Luc. Also cobalt chloride inhibited the TCDD stimulated Cyplal mRNA level as well as EROD activities in both Hepa I and MCF-7 cells. Hypoxic agents such as cobalt chloride, picolinic acid, and desferrioxamine showed inhibition of luciferase activity that was induced by lnM TCDD treatment with dose dependent manner. Concomitant treatment of 150${\mu}$M ferrous sulfate with 1∼100${\mu}$M desferrioxarnine or 1∼100${\mu}$M picolinic acid recovered from the hypoxic agents-inhibited luciferase activity that was stimulated by TCDD. Reciprocally, the hypoxic agents down regulated TCDD induced Cyplal mRNA level and CYP1A1 enzyme activity in Hepa I cells.

• Preventive Nutrition and Food Science
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• 제8권4호
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• pp.365-371
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• 2003

The potential of seven flavonoid glycosides to induce quinone reductase (QR), an anticarcinogenic marker enzyme, in murine hepatoma cells (hepalc1c7) and its mutant cells (BPRc1) was evaluated. Among test compounds, kaempferol-3-O-glucoside, luteolin-6-c-glucoside, and quercetin-3-O-glucoside (Q-3-G) induced QR in hepalc1c7 cells in a dose-dependent manner. However, in BPRc1 cells lacking arylhydrocarbon receptor nuclear translocator (ARNT), only Q-3-G caused a significant induction of quinone reductase at the concentration range of 0.5 to 8 ug/mL, suggesting that it is a monofunctional inducer. Q-3-G induced not only phase 2 enzymes, including QR and glutathione-S-transferase, but also nitroblue tetrazolium reduction activity in HL-60 cells, a biochemical marker for cell differentiation promoting agents. In conclusion, Q-3-G merits further study to evaluate its cancer chemopreventive potential.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.107-107
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• 1997

Cytochrome P450 enzymes have been intensively investigated in hepatic tissues and several mammalian cell lines. Compared to most studies about cytochrome P450 isozymes in liver in vivo and hepatic, cell lines in vitro, the study of cytochrome P450IA1 in human breast cancer cells could be very important to understand the mechanism of the regulation of CYPIA1 gene expression and cell growth. MCF-7 human breast cancer cells are well characterized to study estrogen and antiestrogen action due to the fact that they contain high level of estrogen receptor and have biological markers characterized. And also MCF-7 cells express high level of arylhydrocarbon hydroxylase activity and human cytochrome P450IA1 cDNA was cloned from MCF-7 cells. Ah receptor was characterized in many breast cancer cell lines and polycyclic aromatic hydrocarbon such as 3-MC induced the expression of CYPIA1 gene and cytochrome P450- dependent monooxygenase activity. We undertook a study to examine the effect of estrogens and other chemicals on the regulation of human CYPIA1 gene expression in MCF-7 cells via RTPCR analysis, that might help us to understand the mechanism of the regulation of CYPIA1 gene expression and MCF-7 cell growth. Expression vector containing the functional 5'-regulatory region of human CYPIA1 fused to the CAT reporter gene was transfected into estrogen receptor positive MCF-T cells or estrogen receptor negative MDA-MB-231 cells. After these cells were treated with various chemicals, RTPCR was carried out to measure both CYPIA1 mRNA and CAT mRNA levels. 1nM 3-MC increased in both P450 and CAT mRNA levels over those of control by two folds in MCF-7 cells but does not in MDA-MB-231 cells. Estrogen or tamoxifen or retinoic acid or chrysin decreased in both P450 and CAT mRNA levels that were induced by 3-MC in MCF-7 when each chemical was administered with 3-MC concomitantly. These results suggested that the level of CYPIA1 gene expression is modulated with estrogen-related molecules and make it possible to speculate that ER is related to CYPIA1 gene expression and cell growth in breast cancer cells. [Supported by grants from the Korean Ministry of Education ]

• Archives of Pharmacal Research
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• 제20권5호
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• pp.404-409
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• 1997

Trout CYP1A-CAT expression construct was generated by cloning -3.5 Kb $5^I$ flanking DNA of trout liver CYP1A gene in front of CAT gene at pCAT-basic vector. Hepa 1 cells, which are known to contain a functional arylhydrbcarbon $receptor^I$ were transfected with trout CYP1A-CAT using lipofectin. 3-Methylcholanthrene (1 nM) was added into hepa 1 cells in culture in order to examine if $5^I$ flanking DNA of trout CYP1A gene could interact with mouse transactivating factors to bring about transcription of the chloramphenicol acetyltransferase(CAT) reporter gene. The level of CAT protein was measured by CAT ELISA and the level of CAT mRNA was determined by RTPCR. The treatment of 1 nM 3-methylcholanthrene resulted in two fold increases in CAT protein as well as CAT mRNA compared to untreated control hepa 1 cells. These data indicate that arylhydrocarbon receptors of mouse hepa 1 cells are functional to activate exogenously transfected trout CYP1A-CAT construct in terms of both transcription and translation of CAT. We also examined the effect of 3-methylcholanthrene on endogenous cyplal activity in hepa 1 cell. 3-Methylcholanthrene (1 nM) treatment to hepa 1 cells trahsfected with trout CYP1A-CAT construct stimulated the level of cyp1a1 mRNA by two folds and the activity of ethoxyresorufin-O-deethylase by two fold compared to that of control cells. In this study we reported that trout CYP1A-CAT reporter gene expression construct could be expressed by 3-methylcholanthrene treatment in mouse hepa 1 cells. Thus trout CYP1A-CAT could serve as a good model to study the mechanism of regulation of CYP1A1 gene expression.

• 한국환경성돌연변이발암원학회지
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• 제23권1호
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• pp.30-34
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• 2003

Recent industrial society has human widely exposed to PAHs (polynuclear aromatic hydrocarbons) that are comming from the incomplete combustion of organic material as wider spread environmental contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. Our laboratory have been studied the effect of PAHs in the mouse liver hepa 1 cells. In this study, we examined the mouse liver hepa-l cells as a new bioassay system to evaluate bioactivity of PAHs. We have selected 13 PAHs to examine bioassay using cyp1a1-luciferase reporter gene expression system where cyp1a1 1.6 Kb 5flanking region DNA was cloned in front of luciferase reporter gene and this plasmid was transfected into hepa 1 cells transiently. This cells then used for the study to observe the effect of PAHs. We demonstrated that PAHs induced the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase (EROD) activities in a concentration-dependant manner. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD. Acenaphthene, anthracene, fluorine, naphthalene, pyrene, phenanthrene, carbazole were weak responders to cyp1a1 promoter activity stimulation and EROD induction in hepa 1 cells and these chemicals seemed to respond less to EROD than cyp1a1 promoter activity. Benz(a)anthracene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene, and dibenzo(a,h)anthracene showed strong response to cyp1a1 promoter activity stimulation and also EROD induction in hepa 1cells. Results of dose response study suggested that four strong responding PAHs, such as benzo(a)anthracene benzo(k)fluoranthene, chrysene, and dibenzo(a, h)anthracene might be mediated through arylhydrocarbon receptor system in hepa1 cells.

• 생태와환경
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• 제44권4호
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• pp.364-372
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• 2011

The pale chub (Zacco platypus) is generally found in Asian countries, such as Korea, Japan, and China. Nevertheless, very little information exists about the genes involved in the metabolism of xenobiotics in this species. This species is useful in monitoring the environmental impact on various pollutants in freshwater as a sentinel fish species. We cloned the full-length cDNA sequence of xenobiotic metabolizing cytochrome P450 1A (CYP1A) gene from Z. platypus and characterized it. Tissue distribution and timedependent induction of CYP1A were studied by real-time RT-PCR. Induction pattern of CYP1A was studied by exposing the fish to an arylhydrocarbon receptor agonist, ${\beta}$-naphthoflavone (BNF). The liver showed the highest level of expression in basal state as well as BNF- treated fish. However, appreciable levels of expression were also recorded in Gill and kidney and the least level of expression was observed in the eye. The results of the time-course study revealed an induction in the liver, brain, and gills after 6 h and 12 h in most of the tissues. This study provides an insight into the xenobiotics metabolizing system of Z. platypus and offers baseline information for further research related to biomarker, stress, and adaptive response of this ecologically important fish species in the freshwater environment.