Gas chromatography-mass spectrometry analysis of tert.-butyldimethylsilyl derivatives of 2-acetylaminofluorene and metabolites in isolated rat hepatocytes

Ibañez, Diez; Chessebeuf-Padieu, Martine; Nordmann, Patrice; Padieu, P

doi:10.1007/bf00117869

Cited by 4 publications

(2 citation statements)

References 31 publications

(26 reference statements)

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“…Table 1 shows amounts of 2-AAF metabolites which were authenticated and quantified by GC-MS in the cell line which originated form hepatocytes transfected in vitro with LE-pEJ: (i) 2-AF, the deacetylated metabolite of 2-AAF, (ii) N-OH-2-AAF, the proximate carcinogen : Weisburger et al, 1964, (iii) 1-OH-2-AAF and 3-OH-2-AAF, double bond ring hydroxylated metabolites related to the hepatic activation of 2-AAF into N-OH-2-AAF , and (iv) 9-OH-2-AAF resulting from the hydroxylation of the -CH 2-bridge of the fluorene moiety. All together the total metabolites amounted to 70% of l a l b l C lCt those produced by the control culture of primary hepatocytes (Table 1, and Diez Ibanez et al, 1987). In proliferative LE-pEJ transfected cells N-OH-2-AAF level reached 70% of that of control primary hepatocytes (Table 1.)…”

Section: Transfection Of Rats With Le-pej and Le-p L 7hghneomentioning

confidence: 99%

“…The metabolic activation of a procarcinogen is not a tissue specific marker per se. Nevertheless, the liver is the main site for this 2-AAF activation into N-OH-2-AAF and cultured hepatocytes represent, among all types of cultured tissues, the most proficient cellular model for metabolism of 2-AAF into N-OH-2-AAF (McQueen et al, 1986;Diez-Ibanez et al, 1987). Therefore results shown in Table 1 argue strongly in favor of the hepatocyte progeny of the cell line derived form collagenase dissociated liver cells which were transfected in vitro with the Ha-ras EJ allele.…”

Section: Transfection Of Rats With Le-pej and Le-p L 7hghneomentioning

confidence: 99%

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Maintenance of liver function in long term culture of hepatocytes following In Vitro or In Vivo Ha-ras EJ transfection

et al. 1991

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Collagenase isolated rat hepatocytes were transfected with liposome encapsulated pEJ (LE-pEJ), a plasmid carrying the human cellular activated Ha-rasEJ oncogene. A proliferative cell line was cloned from these cells transfected in vitro. It secreted per day 0.87 micrograms albumin and 0.32 microgram transferrin per 10(6) cells, and 11.06 nmol free and conjugated bile acids (BA) per mg protein. Also, it metabolized 2-acetylaminofluorene (2-AAF) into N- and ring-hydroxylated metabolites and 2-aminofluorene at rates of 1.50, 9.73, and 1.98 nmol/mg cell protein/24 hr, respectively. Rats were i.v. injected with both LE-pEJ and LE-p17hGHneo carrying the hGH cDNA gene, and secreted hGH in the plasma which induced the synthesis of anti-hGH antibodies. A cell line was cloned from cultures of primary hepatocytes isolated from the liver of transfected rats. After 2 to 3 months in culture, this cell line secreted per day 18.9 micrograms albumin and 11.0 micrograms transferrin per 10(6) cells, 38.75 nmol total BA per mg cell protein, and up to 31 ng hGH per 10(6) cells without cloning hGH recombinant cells. A 24 hr control culture of primary hepatocytes isolated from non transfected rats secreted 25.5 micrograms albumin and 11.7 micrograms transferrin per 10(6) cells, and produced 21.64 nmol total BA and 2.13 nmol N-OH-2-AAF per mg cell protein. Hence, Ha-rasEJ transfection of either hepatocytes in vitro or liver cells in vivo, initiated cell cycles leading to presumptive proliferating hepatocytes which express liver function.

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Section: Transfection Of Rats With Le-pej and Le-p L 7hghneomentioning

confidence: 99%

Section: Transfection Of Rats With Le-pej and Le-p L 7hghneomentioning

confidence: 99%

Maintenance of liver function in long term culture of hepatocytes following In Vitro or In Vivo Ha-ras EJ transfection

et al. 1991

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Toxic effects of 70-hydroxycholesterol on rat liver primary cultures, epithelial lines and co-cultures.

et al. 1989

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The toxic effects of 7 beta-hydroxycholesterol (7 beta-OHC) on cultures and co-cultures of rat hepatocytes, rat liver epithelial cell lines, and rat liver fibroblast lines were investigated. Hepatocytes in primary culture or co-cultured with proliferative epithelial cells, were not affected by the presence of 7 beta-OHC at a concentration of 400 microM over a period of 72 hours. In contrast, proliferative cultures of liver epithelial cell lines and liver fibroblast lines were killed by 50 microM 7 beta-OHC within the first 24 hours. Established liver epithelial cells (hyperploid) were more sensitive to 7 beta-OHC than the same line at early passages (diploid). When hepatocytes and liver epithelial cells were co-cultured and treated with 100 microM 7 beta-OHC, only epithelial cells were lysed. A concentration of 50 microM 7 beta-OHC was toxic to co-cultures of liver epithelial cell and fibroblasts together. In a serum-free medium, the cytotoxic concentration of 7 beta-OHC was lower than that in the serum-supplemented medium. Thus, liver epithelial cells cultured alone or co-cultured with hepatocytes were killed at 12.5 microM and 50 microM 7 beta-OHC, respectively. Finally, cholesterol concentrations four-fold that of 7 beta-OHC antagonized the lethal effects of 7 beta-OHC in the serum-free medium.

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Gas chromatographic—mass spectrometric study of deacetylation and oxidation of 2-acetylaminofluorene by rat liver epithelial cell lines upon cocarcinogen induction

Ibañez¹,

Chessebeuf-Padieu²,

Padieu³

1989

Journal of Chromatography B: Biomedical Sciences and Applicatio

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Gas chromatography-mass spectrometry analysis of tert.-butyldimethylsilyl derivatives of 2-acetylaminofluorene and metabolites in isolated rat hepatocytes

Cited by 4 publications

References 31 publications

Maintenance of liver function in long term culture of hepatocytes following In Vitro or In Vivo Ha-ras EJ transfection

Maintenance of liver function in long term culture of hepatocytes following In Vitro or In Vivo Ha-ras EJ transfection

Toxic effects of 70-hydroxycholesterol on rat liver primary cultures, epithelial lines and co-cultures.

Gas chromatographic—mass spectrometric study of deacetylation and oxidation of 2-acetylaminofluorene by rat liver epithelial cell lines upon cocarcinogen induction

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