• Title/Summary/Keyword: 7-ethoxyresorufin-O-deethylase

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Effect of Ethanol Extract from Salvia miltiorrhiza on Chemoprevention and Metastasis of Breast Cancer (단삼 에탄올추출물이 유방암 예방 및 전이에 미치는 영향)

  • Shon, Yun-Hee;Cho, Hyun-Jung;Kim, Mee-Kyung;Jung, Eun-Jung;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.38 no.1
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    • pp.62-66
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    • 2007
  • Ethanol extract from Salvia miltiorrhiza (SME) was tested for breast cancer chemoprevention and metastasis by measuring the activites of cytochrome P45O 1A1, aromatase, ornithine decarboxylase (ODC), and matrix metalloproteinase (MMP)-9. SME significantly inhibited cytochrome P45O 1A1-mediated ethoxyresorufin O-deethylase (EROD) activity in a dose-dependent manner in a concentration range of 100${\sim}$l,200 ${\mu}g/ml$ (p<0.01). Microsomal aromatase (estrogen synthase) activity was suppressed 54.9%${\sim}$77.5% by the SME at concentration of 600${\sim}$l,200 ${\mu}g/ml$. ODC activity induced by 12-O-tet-radecanoylphorbol-13-acetate (TPA) was significantly reduced by the SME 900 and 1,200 ${\mu}g/ml$ (p<0.05) in MCF-7 breast cancer cells. In addition, SME (900 and 1,200 ${\mu}g/ml$) markedly inhibited MMP-9 activity, a key role in cancer metastasis. Therefore, SME is worth further investigation with respect to breast cancer chemoprevention or therapy.

Gender Differences in Activity and Induction of Hepatic Microsomal Cytochrome P-450 by 1-Bromopropane in Sprague-Dawley Rats

  • Kim, Ki-Woong;Kim, Hyeon-Yong;Park, Sang-Shin;Jeong, Hyo-Seok;Park, Sang-Hoi;Lee, Jun-Yeon;Jeong, Jae-Hwang;Moon, Young-Hahn
    • BMB Reports
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    • v.32 no.3
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    • pp.232-238
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    • 1999
  • Sex differences in the induction of microsomal cytochrome P-450 (CYP) and the activities of several related enzymes of Sprague-Dawley rats treated with 1-bromopropane (1-BrP) were investigated. Male and female rats were exposed to 50, 300, and 1800 ppm of 1-BrP per kg body weight (6 h a day,S days a week, 8 weeks) by inhalation. The mean body weight of 1-BrP treated groups increased according to the day elapsed, but four and five weeks respectively after the start of the exposure, the mean body weight of male and female rats had significantly reduced in the group treated with 1800 ppm 1-BrP compared with the control group (p<0.01). While the relative weights of liver increased in both sexes, statistical significance in both sexes was found only in the group receiving 1800 ppm/kg of 1-BrP (p<0.01). The total contents of CYP, $b_5$, NADPH-P-450 reductase, NADH $b_5$ reductase, ethoxyresorufin-O-deethylase (EROD), pentoxyresorufin-O-dealkylase (PROD), and p-nitrophenol hydroxylase (pNPH) activities were examined for the possible effects of 1-BrP. No significant changes in the CYP and $b_5$ contents, NADPH-P-450 reuctase, NADH $b_5$ reductase, ethoxyresorufin-O-deethylase (EROD), and pentoxyresorufin- O-dealkylase (PROD) were observed between the control and treated groups. The activity of pNPH increased steadily with the increase in the concentration of 1-BrP in both sexes, but was significantly increased only in the 1800 ppm-treated group of male rats (p<0.05). When Western blottings were carried out with three monoclonal antibodies (MAb 1-7-1, MAb 2-66-3, and MAb 1-98-1) which were specific against CYP1A1/2, CYP2B1/2, and CYP2E1, respectively, a strong signal corresponding to CYP2E1 was observed in microsomes obtained from rats treated with 1-BrP. Glutathione S-transferase (GST) activity and the content of lipid peroxide significantly increased in the treated groups compared with the control group (p<0.05). These results suggest that 1-BrP can primarily induce CYP2E1 as the major form and that GST phase II enzymes play important roles in 1-BrP metabolism, showing sex-dependence in the metabolic mechanism of 1-BrP in the rat liver.

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Effects on Biomarkers and Endocrine in Muddy Loach (Misgurnus anguillicaudatus) under 21 day Exposure to Methomyl (21일간 methomyl에 노출한 미꾸리의 생물지표 및 내분비계 영향)

  • Han, Sun-Young;Kim, Ja-Hyun;Gwon, Ga-Young;Yeom, Dong-Hyuk
    • The Korean Journal of Pesticide Science
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    • v.16 no.1
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    • pp.69-77
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    • 2012
  • To evaluate the effect of endocrine disruption chemicals (EDCs) to aquatic organisms, muddy loach (Misgurnus anguillicaudatus) was exposed to low concentration methomyl for 21 days in order to identify the effect of biomarkers and endocrine. Vitellogenin (VTG) in blood plasma, which used widely as validated biomarker for endocrine disruption, was significantly greater in male fish exposed to 0.4 mg/L and 2 mg/L methomyl, and in female fish exposed to 0.08 mg/L, 0.4 mg/L, and 2 mg/L methomyl for 21 days (p<0.05). This results suggest that methomyl have probability of endocrine disruption to organism on aquatic system. While inhibition of Acetylcholinesterase (AChE) activity and increase of DNA damage in comet assay were verified by fish exposed to methomyl, change of ethoxyresorufin-O-deethylase (EROD) activity was not occurred, comparing the control group (p<0.05). Indicators at the level of organism such as condition factor (CF), hepato-somatic index (HSI), and gonado-somatic index (GSI) were not influenced by exposure of methomyl. In conclusion, these results showed the possibility of methomyl in regard to not only endocrine disruption but also impacts on biochemical biomarkers to aquatic organisms.

Comparative Study of TCDD Effect on CYP1A1 Gene Expression in Trout and Mouse Liver Cells (송어 간세포와 생쥐 간세포에서 TCDD가 CYP1A1 유전자 발현에 미치는 영향 비교 연구)

  • Kim, Ji-Sun;Min, Kyung-Nan;Sheen, Yhun-Yhong
    • Environmental Mutagens and Carcinogens
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    • v.24 no.3
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    • pp.143-150
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    • 2004
  • In mammalian, cytochrome P4501A1 (CYP1A1) is very important for metabolism of xenobiotics such as PAHs(Polycyclic aromatic hydrocarbon) and heterocyclic amine, and it is induced by environmental contaminants such as PAHs, TCDD(2,3,7,8-tetrchlorodibenzo-p-dioxin) and 3-MC (3-methylcholanthrene). In fish, like mammalian, when it is exposed to environmental contaminants, they cause specific and sensitive induction of CYP1A. Therefore, induction of CYP1A in fish and mammalian is widely used as a biomarker for exposure of environmental contaminants. In this study, to compare the function of Cyp1a1 in fish with it in mammalian, we have used rainbow trout(Oncorhynchys mykiss) hepatoma cells (RTH-149) and mouse hepatocyte (Hepa-I). in order to examine induction of Cyp1a1 by TCDD, we have used the bioassay system. We examined effects of TCDD on the Cyp1a1-luciferase reporter gene activity, 7-ethoxyresorufin O-deethylase(EROD) activity and Cypa mRNA level.

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Cell Culture Microbioassay for the Water Pollution Monitoring (세포배양 생화학적 기법에 의한 수환경오염 평가)

  • 오승민;정규혁
    • Toxicological Research
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    • v.16 no.4
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    • pp.285-291
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    • 2000
  • So far, investigation of environmental pollution has been achieved in field study. This remains the most exhaustive approach, current dimensions of environmental researches and their inherent complexity require that relatively inexpensive and simple laboratory procedures are developed to make possible the screening of large numbers of sites and samples. At this point. microbioassay has been high-lighted. The purpose of this study is to evaluate the water pollution using microbioassay. Two microbioassay methods were optimized and validated for the sensitive and quantitative determination of total toxic effects in the water. EROD(Ethoxyresorufin-O-deethylase) microbioassay was focused to detect PARs, PCBs and dioxinlike components in the water and E-screen assay to xenoestrogens. The EROD microbioassay was executed in rat hepatoma cell line, H4IIE and E-screen assay in MCF7-BUS cell line. Kumho river was selected for this study. 5ι of river water was extracted using combined solid-phase extraction in static adsorption mode with soxhlet extraction. Pollutants adsorbed to the XAD-4 resin were recovered by elution with ethyl acetate and methylene chloride (1 : 9). Toxic effects of extracts were determined by EROD-microbioassay and E-screen assay. EROD activities of water samples were 7.24-72.24 ng/ι MEQ. The estrogenic effect of various water samples is quantitatively evaluated by EEQ. The EEQ of samples range from 0.05 to 6.07 ng-EEQ/ι. These results suggested that Kumho river was highly polluted with organic toxic chemicals.

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Effect of PAH on CYP1A1 Gene Expression in Trout RTH-149 Cells (PAH가 송어 RTH-149세포에서 CYP1A1 유전자 발현에 미치는 영향)

  • Kim, Ji-Sun;Sheen, Yhun-Yhong
    • Environmental Mutagens and Carcinogens
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    • v.24 no.4
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    • pp.155-162
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    • 2004
  • In mammalian, cytochrome P4501A1 (CYP1A1) is very important for metabolism of xenobiotics such as PAHs(Polycyclic aromatic hydrocarbon) and heterocyclic amine, and it is induced by environmental contaminants such as PAHs, TCDD(2,3,7,8-tetrchlorodibenzo-p-dioxin) and 3-MC (3-methylcholanthrene). In fish, like mammalian, when it is exposed to environmental contaminants, they cause specific and sensitive induction of CYP1A. Therefore, induction of CYP1A in fish and mammalian is widely used as a biomarker for exposure of environmental contaminants. In this study, to compare the function of Cyp1a1 in fish with it in mammalian, we have used rainbow trout(Oncorhynchys mykiss) hepatoma cells (RTH-149) and mouse hepatocyte (Hepa-I). in order to examine induction of Cyp1a1 by TCDD, we have used the bioassay system. We examined effects of TCDD on the Cyp1a1-luciferase reporter gene activity, 7-ethoxyresorufin O-deethylase(EROD) activity and Cypa mRNA level.

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Induction of Imposex in Rock Shell, Thais clavigera, Exposed to Organotins and Other Endocrine Disrupting Chemicals

  • Shim, Won Joon
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2003.11a
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    • pp.7-11
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    • 2003
  • Interactions of both feminizing and masculinizing chemicals inducing imposex in gastropod were investigated with a long-term exposure experiment. Imposex-free rock shell(Thais-clavigera)was exposed to TBT and other antagonistic chemicals (nonylphenol and DDT), P450 inducer (phenobarbital) and natural sex hormone (estrogen) mixtures for 60 days. None of imposexed-females were found in control, sham (ethanol) and testosterone exposure group, on the other hand imposex was induced in the TBT and all the TBT+antagonist mixture groups. After 60-day exposure, frequency of imposexed-female ranged from 31-68%. Female mean penis length (FMPL) of imposexed-female ranged from 0.87-2.58 mm, and relative penis length indices were from 7.2 to 21.2%. However, the degrees of imposex were different among the exposure groups. The FMPLS of three exposure groups (TBT+estrogen, + DDT and +nonylphe- not) were less than that of the TBT exposure group. Ethoxyresorufin-O-deethylase (EROD) activities in gastropod decreased in the TBT and all the TBT+antagonist ekposure groups compared to the control group. A negative correlation was abtained between TBT body residue and EROD activity, while a positive relationship was obtained between 737 body residue and the degree of imposex except for the TBT +DDT exposure group. Although 737 concentration was relatively high in the TBT+DDT exposure group, the TBT +DDT exposure group demonstrated low EROD activity and low degree of imposex. These results indicate that a certain antagonistic effect of DDT occurred in induction of imposex by TBT.

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Determination of Dioxin-like Components in the School Waste Incinerator Residues by EROD-microbioassay (EROD-microbioassay에 의한 학교 소각로 잔재 중 다이옥신 유사물질의 측정)

  • 정규혁;오승민;윤완진
    • Journal of Environmental Health Sciences
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    • v.26 no.3
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    • pp.11-17
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    • 2000
  • There are among the most relevant toxic emissions from incinerators such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), dioxin-like polychlorinated biphenyls (PCBs). Induction of cytochrome P4501A1 catalyzed 7-ethoxyresorufin O-deethylase(EROD) activity in mammalian cell culture(EROD bioassay) is thought to be a selective and sensitive parameter used for the quantification of dioxin-like components. In this study, the toxic emissions from several school waste incinerators were evaluated by determination of CYPIA catalytic activity and cytotoxicity using cell culture microbioassay. The incinerator residue and soil samples were collected from the schools located in Kyunggi province from April to June 1999. The samples were extracted in a Soxhlet apparatus using toluene for 20 hours. In order to clean-up, concentrated crude extracts were applied to basic alumina column. The EROD activities of extracts in the H4IIE cells were from 1.91$\pm$0.32 ng-TEQ/g to 24.54$\pm$3.48 ng-TEQ/g of biochemical-TEQ value. In soil samples, CYP1A catalytic activity was 0.09~0.64 ng-TEQ/g. EROD bioassay, seems to be a useful short-term bioassay when information about the biological response of complex environmental samples is needed. Although further study is needed, these results indicate that the potent toxic emissions are produced from school waste semi-incinerators.

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Studies on Toxicological Evaluation of Freshwater Sediment using a PLHC-1 Cell Comet Assay (PLHC-1세포주의 Comet assay를 이용한 하천 퇴적토의 생태독성평가)

  • Bak, Jeong-Ah;Hwang, In-Young;Baek, Seung-Hong;Kim, Young-Sug
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.23-30
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    • 2011
  • In this study, the Comet assay (evaluation of DNA damage) used the fish hepatocellular carinoma cell, PLHC-1, was tried to the sediment extract obtained from freshwater to understand its applicability as a tool for monitoring sediment toxicity. In parallel, induced EROD (7-ethoxyresorufin- O-deethylase) activity and DNA damage (TEM values) in PLHC-1 cells were measured for establishing the tandem endpoints of the PLHC-1cell test to test the ecotoxicity of sediment. Among several study sites in a small river passed through downtown and industrial park area, one of them, site B, showed a higher level of EROD activity and DNA damage than other sites. It indicates that a tandem endpoints of PLHC-1 cells could be useful tools for assessing the toxicity of sediment. The sensitivity of Comet assay with PLHC-1 cells was a little higher than that with a blood cell of frog tadpoles to the solvent extract of sediment. According to the results, a PLHC-1 cell-Comet assay could be used as a useful tool for evaluating ecotoxicity of the freshwater sediment. In addition, more detailed studies are needed to the contaminated site.

Effects of Trialkyltin in vitro on the Microsomal Monooxygenase System of Digestive Gland in the Clam, Coelomactra antiquata (유기주석화합물이 명주조개 (Coelomactra antiquata)의 약물대사효소계에 미치는 영향)

  • Jeon Joong Kyun;Lee Mee Hee;Kim Do Jin;Shim Won Joon;Oh Jae Ryong;Lee Soo Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.185-190
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    • 2002
  • This study was carried out to measure the in vitro interaction of trialkyltin with the microsomal monooxygenase (MFO) system of the clam, Coelomactra antiquata. Cytochrome P450 (CYP) level and 7-ethonvesorufin-O-deethylase (EROD) activity were invetigated in the microsome isolated from digestive gland of the clam (C. antiquata) exposed to tributyltin chloride (TBTC), bis-tributyltinoxide (TBTO) and triphenyltin chloride (TPTC). The specific contents of CYP in clam microsome exposedto 0.4 mM TBTC, TBTO and TPTC for 20 minutes were decreased 52, 72 and $40\%$, respectively, compared to control group. The EROD activities also were inhibited by exposure to TBTO ($92\%$) and TPTC ($85\%$) except for TBTC, The level of CYP and the EROD actintles were decreased according to the OTC exposure concentrations. The toxic effects on the level of CYP and the EROD activities were in order of TPTC>TBTC>TBTO in this study. The measurement of CYP level and EROD activity could be applied as a biomarker for environmental study.