Cytochrome P4501A induction and inhibition by 3,3′,4,4′-tetrachlorobiphenyl in an Ah receptor-containing fish hepatoma cell line (PLHC-1)

Hahn, Mark E.; Lamb, Teresa M.; Schultz, Mary E.; Smolowitz, Roxanna; Stegeman, John J.

doi:10.1016/0166-445x(93)90030-5

Cited by 245 publications

(159 citation statements)

References 59 publications

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“…Several P450 and GST catalytic activities were assayed as a means to determine functionality of P450 and GST proteins encoded by the measured mRNA. Specifically, CYP1A1-dependent ethoxyresorufin O-deethylase (EROD) activity in HSC S9 fractions was measured using a fluorescent microplate reader (Hahn et al, 1993). To determine the functionality of CYP3A5 and CYP3A7 mRNA expression, human fetal HSC were incubated with 12 M midazolam for 6 h in a shaking water bath at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Cytochrome P450 and GlutathioneS-Transferase mRNA Expression in Human Fetal Liver Hematopoietic Stem Cells

et al. 2006

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ABSTRACT:During fetal development, the liver serves as the primary hematopoietic organ in which hematopoietic stem cells (HSC) capable of initiating long-term hematopoiesis comprise a large proportion of the hepatic cell population. Although HSC are potential targets for transplacental chemicals, little is known regarding their xenobiotic biotransformation ability. We quantitated the steady-state mRNA expression of six cytochrome P450 (P450) and 11 glutathione Stransferase (GST) isoforms in CD34؉ -selected HSC isolated from second trimester human fetal liver donors, genotyped donors for polymorphic hGSTM1 and hGSTT1 status, and analyzed gene expression in HSC relative to total liver from donors of similar gestational ages. Several P450 isoforms, including CYP1A1, CYP2E1, CYP3A4, and CYP3A5, were expressed at low levels in HSC (relative mRNA expression CYP3A5 > CYP1A1 > CYP2E1 > CYP3A4).CYP1A2 and CYP3A7 were not detected in HSC. The CYP3A4/5 mRNA expression in HSC was accompanied by detectable CYP3A protein and low midazolam oxidation activity. Several GST isoforms, including hGSTM1, hGSTM2, hGSTM4, and hGSTP1, were significantly higher in HSC as compared with total fetal liver. With the exception of hGSTA4, alpha class GST were not detected in HSC. GST expression in HSC was accompanied by substantial GST catalytic activity toward 1-chloro-2,4-dinitrobenzene. In summary, our data indicate that fetal liver CD34 ؉ -derived HSC constitutively express several P450 isoforms at low levels relative to total hepatic cell populations but have a higher capacity for GST conjugation reactions through mu and pi class isoforms. The functional ramifications of these observations are discussed relative to the sensitivity of human fetal HSC to transplacental chemical injury.

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Section: Methodsmentioning

confidence: 99%

Cytochrome P450 and GlutathioneS-Transferase mRNA Expression in Human Fetal Liver Hematopoietic Stem Cells

et al. 2006

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“…EROD activity: CYP1A1 activity was determined using a fluorometric, kinetiô assay for EROD activity Hahn et al, 1993. Assays were run in 48 well plates with 2 uM 7-ethoxyresorufin and 1.0 mM NADPH final concentrations.…”

Section: Methodsmentioning

confidence: 99%

Characterization of resistance to halogenated aromatic hydrocarbons in a population of Fundulus heteroclitus from a marine superfund site

Bello¹

1999

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New Bedford Harbor NBH, MA, is contaminated with halogenated aromatic hydrocarbons HAH including some potent aryl hydrocarbon receptor AhR agonists. To determine ifFundulus heterocitus from NBH have developed resistance to HAll, we examined the inducibility of cytochrome P4501A1 CYP1A1 in fish from NBH and Scorton Creek SC, reference site. Despite higher PCB concentrations in NBH than in SC fish --1 500-fold -CYP1A1 expression, in most tissues, was not higher in NBH fish than in SC fish. Glutathione S-transferase GST and UDP-glucuronosyltransferase UGT activities were higher in NBH fish than in SC fish, but only when fish were collected during different seasons. GST activity was higher in the intestines ofNBH fish than in any other tissue.2,3,7,8-Tetrachlorodibenzoftiran TCDF induced CYP1A1 expression, in all tissues examined, in SC fish. In contrast, NBH fish showed little CYP1A1 induction by any measure, in any tissue. Hepatic GST activity was induced only in male NBH fish. Hepatic UGT activity showed no relationship to treatment in fish from either site.2,3,7,8-Tetrachlorodibenzo-p-dioxin TCDD and J3-naphthoflavone BNF induced CYP1A1 activity to the same level in primary cultures ofhepatocytes from either SC orNBH fish. However, hepatocytes from NBH fish were 14-fold less sensitive to TCDD and 3-fold less sensitive to BNF than hepatocytes from SC fish.To examine the heritability ofresistance, NBH and SC F1 fish were treated with 3H-TCDD or BNF. 3H-TCDD induced CYP1A1 expression only in SC F1 fish. BNF induced CYP1A1 expression in both SC and NBH F1 fish. There was no significant difference in hepatic 3H-TCDD concentrations between SC and NBH F1 fish.Hepatic AhR content, as measured by photoaffinity labeling with '25I-N3Br2DD, was lower in NBH fish than in SC fish and lower in males than in females. After 90 days in captivity, the sex difference persisted, but the site difference did not. TCDF induced hepatic AIIR content in NBH F1 fish.These results indicate that NBH Fundulus have developed a pre-translational, systemic, heritable resistance to HAHs. These findings suggest that an alteration in the AIIR pathway is responsible for this resistance; this is the subject ofcontinuing research. Next, I owe more than I can say to my thesis advisor and friend Mark Hahn. He was always available whenever I needed to consult on ideas, was endlessly tolerant of the bizarre hours I kept, and fostered a truly enjoyable work environment.Bruce Woodin was an ever accommodating and delightful person to work with. I want to thank him for all ofhis help with dissections, his enlightening field trips, and for generally making my time here that much more enjoyable. I thank Diana Franks for her work with the primary culturing of the Fundulus hepatocytes, for her help with fish care and for generally helping to make my life a little easier. Diane Nacci, Ken Rocha, and the EPA laboratory in Narragansette for kindly providing fish. Diane also for her help during the planning stages ofthis work and her willingness to exchange r...

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“…The presence of CYP1A was detected by immunoblot analysis of microsomal preparations as described previously with slight modification (Hahn et al,1993;Stegeman et al,1995). Microsomal protein (30 to 70 µg) and a range of standards of CYP1A from scup (Stenotomus chrysops) were resolved on a 4-20 % acrylamide gradient gel in TRIS-glycine with SDS (Jule,Inc, Milford, CT).…”

Section: Immunoblotting For Cyp 1amentioning

confidence: 99%

“…Induction of fish CYP1A by AhR agonists have been studied in highly exposed natural populations, in laboratory studies of fish taken from the field, in fish embryos and in fish hepatoma cells (Hahn et al,1993;Guiney et al,1997;Schlezinger and Stegeman,2000b). With the establishment of methods to maintain and subculture teleost fish endothelial cells (Garrick,2000) we can now address questions about effects of xenobiotics on the endothelium that could not be tested previously.…”

Section: Introductionmentioning

confidence: 99%

Cytochrome P4501a Induced Differentially in Endothelial Cells Cultured From Different Organs of Anguilla Rostrata

Garrick

Woodin

Stegeman

2005

In Vitro Cell Dev Biol Anim

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Abstract:Endothelial cells are a structural barrier and an active regulator of many bodily processes. CYP1A activity is induced in the endothelium of teleosts and mammals exposed to lipophilic xenobiotics, such as polycyclic aromatic hydrocarbons, and can have significant consequences for endothelial functions. We exposed cultures of characterized endothelial cells from the heart, kidney and rete mirabile of the eel, Anguilla rostrata, to AhR agonists. In heart endothelial cells the maximum response (based on EROD activity) to TCDD, 113 pmol/mg-min, was at 1 nM TCDD and the peak response to βNF, 135 pmol/mg-min, was at 3 µM βNF. The maximum response to

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Cytochrome P4501A induction and inhibition by 3,3′,4,4′-tetrachlorobiphenyl in an Ah receptor-containing fish hepatoma cell line (PLHC-1)

Cited by 245 publications

References 59 publications

Cytochrome P450 and GlutathioneS-Transferase mRNA Expression in Human Fetal Liver Hematopoietic Stem Cells

Cytochrome P450 and GlutathioneS-Transferase mRNA Expression in Human Fetal Liver Hematopoietic Stem Cells

Characterization of resistance to halogenated aromatic hydrocarbons in a population of Fundulus heteroclitus from a marine superfund site

Cytochrome P4501a Induced Differentially in Endothelial Cells Cultured From Different Organs of Anguilla Rostrata

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