Utilization of an established rat hepatoma cell line for mutation studies

Green, Howard A.; Kram, David; Williams, Jerry R.

doi:10.1016/0165-1161(82)90031-0

Cited by 4 publications

(2 citation statements)

References 11 publications

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“…We have previously described a mutagenesis assay utilizing the rat hepatoma cell line, H4IIE (Green et al, 1982). The two cell lines have a number of common characteristics: l) both appear capable of metabolizing P450 and P44s-dependent promutagens; 2) both show optimal seeding densities of approximately 2 X 105 cells per 100 mm plate for the selection of mutants; 3) both can be used for cytogenetic studies (Dean et al, 1980(Dean et al, , 1983; and 4) both show similar phenotypic expression periods of 8 to 10 days.…”

Section: Discussionmentioning

confidence: 99%

“…Detecting forward mutations at specific loci in cultured mammalian cells has proven a useful tool in evaluating the potential hazard for genetic damage of chemicals and radiations (Chu and Mailing, 1968;Arlett, 1977;Green et al, 1982). Simulating the in vivo biotransformation of promutagens/procarcinogens to an active genotoxic form, however, poses a major problem for such in vitro assays.…”

Section: Introductionmentioning

confidence: 99%

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Use of an established human hepatoma cell line with endogenous bioactivation for gene mutation studies

1988

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Genetic toxicology assays that rely on S9 microsomal mixes are subject to artifacts related to the generation of mutagenic metabolites by acidic pHs, variation in individual isolations of microsomes and the failure of subcellular fractions to faithfully produce metabolites generated in intact cells. We have developed a gene mutation assay utilizing the human hepatoma cell line HepG2, which has been shown to metabolize a broad spectrum of promutagens. Optimal conditions for assaying the induction of 6-thioguanine-resistant mutants in this cell line include: 1) growth of colonies for three weeks on lethally irradiated feeder layers of 10(6) thioguanine-resistant HepG2 cells (average plating efficiency = 60-80%); 2) a thioguanine concentration in selection dishes of 10(-4) M with a maximum seeding density of 2.5 x 10(5) cells per 100 mm culture dish; and 3) a minimum expression time of 6 days. In addition to ultraviolet light C (254 nm), a cytochrome P450 (cyclophosphamide)-dependent and a cytochrome P448 (aflatoxin B1)-dependent promutagen were shown to induce cytotoxicity and mutations in this test system. The present studies, therefore, suggest that the HepG2 cell line may be useful for a variety of assays in genetic toxicology.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Use of an established human hepatoma cell line with endogenous bioactivation for gene mutation studies

1988

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Induction of cytochrome(s) P₄₅₀‐dependent drug metabolism in cultured MH₁C₁ hepatoma cells

Ferro

Bassi

Marinari

et al. 1984

Cell Biochemistry & Function

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A cell line derived from a Morris hepatoma, MH1C1, was examined for its in vitro expression of monooxygenases. These cells were found to contain different forms of cytochrome P450, as shown by the response to inducers, namely phenobarbital (PB), 3-methylcholanthrene (MC) and metyrapone (MP). MH1C1 cell monolayers exposed to PB or MC showed an increase in the concentration of two spectrally distinct forms of cytochrome P450. The PB and MC treatments elicited enzyme activities towards the substrates aminopyrine and benzo(a)pyrene, respectively. The cell treatment with metyrapone led to a simultaneous stimulation of aminopyrine demethylase and benzo(a)pyrene hydroxylase activities, so underlining the peculiar features of this inducer.

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Induction of sister chromatid exchanges in human and rat hepatoma cell lines by cyclophosphamide and phosphoramide mustard and the effects of cytochrome P-450 inhibitors

Dearfield

Jacobson‐Kram

Huber

et al. 1986

Biochemical Pharmacology

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Utilization of an established rat hepatoma cell line for mutation studies

Cited by 4 publications

References 11 publications

Use of an established human hepatoma cell line with endogenous bioactivation for gene mutation studies

Use of an established human hepatoma cell line with endogenous bioactivation for gene mutation studies

Induction of cytochrome(s) P₄₅₀‐dependent drug metabolism in cultured MH₁C₁ hepatoma cells

Induction of sister chromatid exchanges in human and rat hepatoma cell lines by cyclophosphamide and phosphoramide mustard and the effects of cytochrome P-450 inhibitors

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Utilization of an established rat hepatoma cell line for mutation studies

Cited by 4 publications

References 11 publications

Use of an established human hepatoma cell line with endogenous bioactivation for gene mutation studies

Use of an established human hepatoma cell line with endogenous bioactivation for gene mutation studies

Induction of cytochrome(s) P450‐dependent drug metabolism in cultured MH1C1 hepatoma cells

Induction of sister chromatid exchanges in human and rat hepatoma cell lines by cyclophosphamide and phosphoramide mustard and the effects of cytochrome P-450 inhibitors

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Induction of cytochrome(s) P₄₅₀‐dependent drug metabolism in cultured MH₁C₁ hepatoma cells