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검색결과 224건 처리시간 0.061초

Human CYP1A2 Promoter Fused-Luciferase Gene Constructs Hardly Respond to Polycyclic Hydrocarbons in Transient Transfection Study in HepG2 Cells

  • Chung, Injae
    • Toxicological Research
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    • 제16권2호
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    • pp.95-100
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    • 2000
  • In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

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Bioassays of Polycyclic Aromatic Hydrocarbons in ZR-75-1 Human Breast Cancer Cells

  • Min, Kyung-N.;Sheen, Yhun-Y.
    • 한국환경성돌연변이발암원학회지
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    • 제23권1호
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    • pp.35-40
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    • 2003
  • Polycyclic aromatic hydrocarbons (PAH) are frequently detected in food, water, soil, and sediment and are widespread environmental pollutants formed by the incomplete combustion of fossil fuels, woods and other organic matter. PAHs are considered to be probable human carcinogens. The mechanism of action of PAHs has been studied extensively, however it is not clear how PAHs turn on CYP1A1 in human breast cancer. Our laboratory have been studied the effect of PAHs in the human breast cancer cell MCF7. In this study, we examined the ZR-75-1 human breast cancer cells as a new system to evaluate bioactivity of PAHs. ZR-75-1 human breast cancer cell line responses to estrogen and progesteron. We have been able to estbilish long term culture system of this cells then used for the study to observe the effect of PAHs. We demonstrate that PAHs induced the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase (EROD) activity in a concentration-dependant manner. RT-PCR analysis indicated that PAHs significantly up-regulate the level of CYP1A1 mRNA. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD. Apparently, ZR-75-1 cells have Aryl hydrocarbon receptors, therefore it would be good experimental tool to study the cross-talk between PAHs and steroid actions.

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HYSTERETIC MODELING ON THE CONVECTIVE TRANSPORT OF ORGANIC SOLVENT IN AN UNSATURATED SOIL ZONE

  • Lee, Kun-Sang
    • Environmental Engineering Research
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    • 제11권5호
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    • pp.241-249
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    • 2006
  • A mathematical model is described for the prediction of convective upward transport of an organic solvent driven by evaporation at the surface, which is known as the major transport mechanism in the in-situ photolysis of a soil contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD). A finite-element model was proposed to incorporate the effects of multiphase flow on the distribution of each fluid, gravity as a driving force, and the use of hysteretic models for more accurate description of k-S-p relations. Extensive numerical calculations were performed to study fluid flow through three types of soils under different water table conditions. Predictions of relative permeability-saturation-pressure (k-S-p) relations and fluids distribution for an illustrative soil indicate that hysteresis effects may be quite substantial. This result emphasizes the need to use hysteretic models in performing flow simulations including reversals of flow paths. Results of additional calculations accounting for hysteresis on the one-dimensional unsaturated soil columns show that gravity affects significantly on the flow of each fluid during gravity drainage, solvent injection, and evaporation, especially for highly permeable soils. The rate and duration of solvent injection also have a profound influence on the fluid saturation profile and the amount of evaporated solvent. Key factors influencing water drainage and solvent evaporation in soils also include hydraulic conductivity and water table configuration.

PAH가 CYP1B1 유전자 발현에 미치는 영향 (Effect of PAH on CYP1B1 Gene Expression)

  • 서미정;민경난;신윤용
    • 한국환경성돌연변이발암원학회지
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    • 제24권3호
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    • pp.121-127
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    • 2004
  • Cytochrome P4501B1(CYP1B1) is known to be inducible by xenobiotic compounds such as policyclic aromatic hydrocarbon(PAH) and dioxins such as 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). And these induction of CYP1B1 is also regulated by many categories of chemicals. In order to investigate the effects of several chemicals on CYP1B1 gene expression in Hepa-I and MCF-7 cells, 5' flanking DNA of human CYP1B1 was cloned into pGL3 basic vector containing luciferase gene, and then transfected into these cells. After treatment of chemicals, the luciferase activity was measured. CYP1B1 enzyme metabolize PAHs and estradiol. CYP1B1 metabolize estradiol to 4-hydrozyestradiol that is considered as carcinogenic metabolite. Luciferase activity was induced about 20 folds over that control by 1 nM TCDD (2,3,7,8-tetrachloto-p-dioxin). Recent industrialized society, human has been widely been exposed to widespread environmental contaminants such as PAHs(polycyclic aromatic hydrocarbon) that are originated from the imcomplete combustion of hydrocarbons. PAHs are known to be ligands of the AhR(aryl hydrocarbon receptor). Induction of cytochrome P4501B1(CYP1B1) in cell culture is widely used as a biomarker for PAHs. Therefore we have studied the effect of PAHs in the human breast cancer cells MCF-7 to evaluate bioactivity of PAHs. We have used the United State of America EPA selected 13 different PAHs, PAHs mixtures and extracts from environmental samples to evaluate the bioassay system. We examined effects of PAHs on the CYP1B1-luciferase reporter gene and CYP1B1 mRNA level. Benzo(k)fluoranthene and dibenzo(a, h)anthracene showed strong response to CYP1B1 promoter activity stimulation, and also CYP1B1 mRNAs increase in MCF-7 cells in a concentration-dependent manner. Acenaphthene, anthracene, benzo(b)fluoranthene, fluorene, fluoranthene, anphthanlene, pyrene, phenanthrene and carbazole were weak responders in MCF-7 cells. RT-PCR analysis indicated that PAHs significantly up-regulate the level of CYP1B1 mRNA.

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