DNA Adduct Formation and Removal in N-Acetoxy-2-Acetylaminofluorene-Exposed Cultured Cells and in Organs from Rats Fed 2-Acetylaminofluorene

Poirier, Miriam C.; Yuspa, Stuart H.; True, B'Ann; Laishes, Brian A.

doi:10.1007/978-1-4684-4400-1_33

Cited by 5 publications

(3 citation statements)

References 6 publications

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“…As can be seen from the effects of PCP (Table 5) and salicylamide (Table 7) on DNA repair synthesis caused by AAF and N-OH-AAF, sulfation of N-OH-AAF is probably involved in the formation of genotoxic metabolites. Although less acetylated than deacetylated arylamine-DNA adducts are found in vivo (Meerman et al, 1981;Poirier et al, 1983), our findings may be explained by the fact that more acetylated-DNA adducts are found in isolated hepatocytes and that acetylated products are removed faster than deacetylated-DNA adducts (Poirier et al, 1983). Further, the sulfate ester is known to be highly reactive and to cause cytotoxic effects judged from in vivo experiments (Irving, 1975), this is supported by results of the present study (Tables 9 and 10).…”

Section: Discussionmentioning

confidence: 99%

Modulation of genotoxic and cytotoxic effects of aromatic amines in monolayers of rat hepatocytes

Holme¹,

Sønderland²

1984

Cell Biol Toxicol

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Cultured rat hepatocytes exposed to 2-acetylaminofluorene (AAF), 2-aminofluorene (AF) or N-hydroxy-2-acetylaminofluorene (N-OH-AFF) for 3 hrs resulted in an increase in DNA repair measured as unscheduled DNA synthesis, with N-OH-AAF greater than AAF greater than AF. Cytotoxic effects were only seen with N-OH-AAF above 10(-6) M. alpha-Naphthoflavone increased the unscheduled DNA synthesis and cytotoxic effects of N-OH-AAF, whereas it decreased DNA repair and the covalent binding of AAF to cellular proteins. In contrast, very little effects of paraoxon were seen on the repair synthesis elicited by AAF, AF or N-OH-AAF. The addition of ascorbate reduced the covalent binding of AAF, the DNA repair synthesis caused by AAF and N-OH-AAF, and the cytotoxic effects of N-OH-AAF. The addition of pentachlorophenol or salicylamide all resulted in similar effects as ascorbate, through reduction of sulfation. Galactosamine, an inhibitor of glucuronidation, and the nucleophile GSH caused no or only minor effects of the activation of AAF, AF or N-OH-AAF as judged from the endpoints tested. These results are consistent with an arylnitrenium ion, a sulfate ester or a free radical as the arylamine metabolite causing cellular DNA damage, whereas the sulfate ester or a radical intermediate may be responsible for the cytotoxic effects of N-OH-AAF.

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Section: Discussionmentioning

confidence: 99%

Modulation of genotoxic and cytotoxic effects of aromatic amines in monolayers of rat hepatocytes

Holme¹,

Sønderland²

1984

Cell Biol Toxicol

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“…Therefore the amount of adduct remaining after 28 days on control diet would appear to be a reflection of the amount of C-8 adduct pre- of C-8 adducts in male WF rats fed 0.02% 2-AAF. Seven animals were fed 0.02% 2-AAF for 25 days, at which time three were sacrificed (A, 25 days), four were fed 0.02% [3H]-2-AAF (219 mCi/mmole) for 3 days, at which time two were sacrificed (A, 28 days) and two were given control diet for a subsequent 28 days (A). Values for total C-8 adduct obtained by RIA from these liver DNAs are represented by A and /v; values for the newly formed (25-28 days) radioactive adducts in the same animals are represented by * and Ol.…”

Section: Removal Of C-8 Adducts From the Livers Of 2-aaf-fed Rats Aftmentioning

confidence: 99%

Determination of 2-acetylaminofluorene adducts by immunoassay

Poirier¹,

True²,

Laishes³

1983

Environ Health Perspect

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Antisera elicited in rabbits were used in radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) to determine femtomole quantities of deoxyguanosin-(8-yl)-acetylaminofluorene (dg-8-AAF) and deoxyguanosin-(8-yl)-aminofluorene (dg-8-AF). These adducts have been monitored in liver and kidney DNA of male Wistar-Furth rats fed 0.02% or 0.04% 2-acetylaminofluorene (2-AAF) either continuously or for a limited time followed by an interval on control diet. After 24 hr of 0.02% 2-AAF feeding, substantial levels of binding (80 fmole/μg DNA) were observed in liver DNA and increased with time, reaching a plateau of approximately 230 fmole/μg DNA at 30 days and thereafter. During the first week of continuous feeding about 80% of the total C-8 adducts in the liver DNA were deacetylated (dG-8-AF). By 25-60 days, dG-8-AF represented 97-100% of all C-8 adducts as measured by RIA and confirmed by HPLC. Values for C-8 adduct formation in kidney DNA were severalfold lower than in liver and dG-8-AF represented >90% of C-8 adducts at all times studied. In removal or repair experiments, rats were fed 2-AAF for 3, 7 or 28 days, the 2-AAF diet was discontinued and the liver adducts assayed after intervals on control diet. When dietary 2-AAF administration was for 3 or 7 days, removal of adducts was efficient and almost complete by 28 days on control diet, with preferential retention of dG-8-AF. However, when dietary 2-AAF administration was for 28 days, adduct levels were higher, the repair capacity was saturated and the removal of C-8 adducts was not complete after control diet for a 28-day interval. In a preliminary experiment when [3H]-2-AAF was fed for 3 days, after 25 days of 0.02% 2-AAF, the rates of newly formed adduct formation and removal were similar to those observed for the initial 3 days of 2-AAF feeding. These results demonstrate the predominance and persistence of dG-8-AF in liver and kidney DNA of 2-AAF-fed rats and suggest that the repair capacity of the whole rat liver was not diminished after 1 month of 2-AAF feeding.

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“…Antiguanosin48-yl)acetylmainofluorene, elicited in rabbits, has been utilized to quantitate carcinogen-DNA adducts by immunological assays and the results compared favorably to other analytical procedures (2,3). The antiserum is specific of the acetyl-*In Vitro Pathogenesis Section, Laboratory of Cellular Carcinogenesis and Tumor Promotion, National Cancer Institute, Bethesda, MD 20205. tAuthor to whom reprint requests should be addressed.…”

Section: Introductionmentioning

confidence: 99%

Determination of 2-Acetylaminofluorene Adducts by Immunoassay

Poirier¹,

True²,

Laishes³

1983

Environmental Health Perspectives

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Antisera elicited in rabbits were used in radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) to determine femtomole quantities of deoxyguanosin-8-yl-acetylaminofluorene (dg-8-AAF) and deoxyguanosin-8-yl-aminofluorene (dg-8-AF). These adducts have been monitored in liver and kidney DNA of male Wistar-Furth rats fed 0.02% or 0.04% 2-acetylaminofluorene (2-AAF) either continuously or for a limited time followed by an interval on control diet. After 24 hr of 0.02% 2-AAF feeding, substantial levels of binding (80 fmole/4g DNA) were observed in liver DNA and increased with time, reaching a plateau of approximately 230 fmole/,Ag DNA at 30 days and thereafter. During the first week of continuous feeding about 80% of the total C-8 adducts in the liver DNA were deacetylated (dG-8-AF). By 25-60 days, dG-8-AF represented 97-100% of all C-8 adducts as measured by RIA and confirmed by HPLC. Values for C-8 adduct formation in kidney DNA were severalfold lower than in liver and dG-8-AF represented >90% of C-8 adducts at all times studied.In removal or repair experiments, rats were fed 2-AAF for 3, 7 or 28 days, the 2-AAF diet was discontinued and the liver adducts assayed after intervals on control diet. When dietary 2-AAF administration was for 3 or 7 days, removal of adducts was efficient and almost complete by 28 days on control diet, with preferential retention of dG-8-AF. However, when dietary 2-AAF administration was for 28 days, adduct levels were higher, the repair capacity was saturated and the removal of C-8 adducts was not complete after control diet for a 28-day interval. In a preliminary experiment when[3H]-2-AAF was fed for 3 days, after 25 days of 0.02% 2-AAF, the rates of newly formed adduct formation and removal were similar to those observed for the initial 3 days of 2-AAF feeding. These results demonstrate the predominance and persistence of dG-8-AF in liver and kidney DNA of 2-AAFfed rats and suggest that the repair capacity of the whole rat liver was not diminished after 1 month of 2-AAF feeding.

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DNA Adduct Formation and Removal in N-Acetoxy-2-Acetylaminofluorene-Exposed Cultured Cells and in Organs from Rats Fed 2-Acetylaminofluorene

Cited by 5 publications

References 6 publications

Modulation of genotoxic and cytotoxic effects of aromatic amines in monolayers of rat hepatocytes

Modulation of genotoxic and cytotoxic effects of aromatic amines in monolayers of rat hepatocytes

Determination of 2-acetylaminofluorene adducts by immunoassay

Determination of 2-Acetylaminofluorene Adducts by Immunoassay

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