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  • 1
    Digitale Medien
    Digitale Medien
    A carcinogen-DNA binding assay as a biomarker screen for identifying potential chemopreventive agents (1997)
    Springer
    Methods in cell science 19 (1997), S. 45-48 
    Details
    ISSN: 1573-0603
    Schlagwort(e): Benzo[a]pyrene ; Carcinogen-DNA binding ; Chemopreventive agents ; Ellagic acid ; Human bronchial epithelial cells
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A biomarker prescreen assay using inhibition of DNA damage induced by the binding of a carcinogen, benzo[a]pyrene (B[a]P) to human bronchial epithelial cells (BEAS-2B) is described. The BEAS-2B cells are immortalized but are essentially normal by being non-tumorigenic in animals and having most of the metabolic functions intact. BEAS-2B cells plated for 18 hours are exposed to a chemopreventive test agent for 2 hours followed by addition of 3H-B[a]P and incubation for an additional 18 hours. DNA is extracted by a non-phenolic procedure and after determining DNA purity and content, the radioactivity is measured. The percent inhibition of radioactivity per mg of DNA sample in the agent-treated samples compared to untreated cell control is calculated. Ellagic acid (15 μM) is used as a positive inhibitor to ensure the assay conditions are appropriate. The assay is easy to perform and provides a reliable screening model for chemopreventive activity using a large number of compounds in a relatively short time. The preincubation with test agents provides an additional advantage to the screening system as it would facilitate the detection of agents that are effective by inhibiting carcinogen-activating enzymes.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1023/A:1009798621899
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  • 2
    Digitale Medien
    Digitale Medien
    Development of glutathione induction as a screening assay for potential chemopreventive agents (1997)
    Springer
    Methods in cell science 19 (1997), S. 53-56 
    Details
    ISSN: 1573-0603
    Schlagwort(e): Buffalo rat liver cells ; Chemoprevention screening ; Glutathione
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A simple biochemical assay for screening potential chemopreventive agents using induction of (reduced) glutathione (GSH), in rat liver (BRL-3A) cells is described. BRL-3A cells, cultured to subconfluency, are exposed to potential chemopre-ventive agents for 24 hours and solubilized by sonication. The cytosolic fraction is then assayed for GSH by reacting with O-phthalaldehyde (OPT) to form a fluorescent product that is activated at 350 nm and has an emission peak at 420 nm. The amount of GSH in samples is determined from a regression curve generated from known concentrations of GSH. The percent induction is estimated by comparing the levels in untreated control (basal level) and the agent-treated groups. N-acetyl-l-cysteine at 10 μM is routinely included in each assay as a positive inducer of GSH.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1023/A:1009702822807
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  • 3
    Digitale Medien
    Digitale Medien
    Induction and prevention of carcinogen-induced precancerous lesions in mouse mammary gland organ culture (1997)
    Springer
    Methods in cell science 19 (1997), S. 19-24 
    Details
    ISSN: 1573-0603
    Schlagwort(e): Chemoprevention ; Mammary alveolar lesions ; Mammary gland ; Organ culture
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Mouse mammary glands respond to growth promoting hormones in organ culture. In the presence of insulin, prolactin, aldostrone, and hydrocortisone, the glands exhibit extensive proliferation within 10 days of culture mimicking the mammary alveolar structures observed during pregnancy. However withdrawal of prolactin and steroids from the medium for an additional 14 days results in the disintegration of the alveolar structures resembling the mammary morphology observed during the involution stage. During the growth promoting phase if the glands are exposed to 7, 12, dimethylbenz(a)anthracene (DMBA) for 24 hours and cultured through the entire 24 days of culture period, they develop precancerous lesions. This model is highly reproducible and extensively utilized to evaluate efficacy of potential chemopreventive agents against carcinogen-induced mammary lesions.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1023/A:1009770720081
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  • 4
    Digitale Medien
    Digitale Medien
    Induction of NADPH-quinone reductase as a screening assay for potential chemopreventive agents (1997)
    Springer
    Methods in cell science 19 (1997), S. 63-66 
    Details
    ISSN: 1573-0603
    Schlagwort(e): Chemoprevention ; Human liver cells ; Multi-well assay ; Oltipraz ; Quinone reductase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract An in vitro biochemical assay using induction of a phase II enzyme, NADPH-quinone reductase (NADPH-QR) for selection and identification of potential chemopreventive agents is described. A normal human liver cell line (Chang liver cells) or a rodent rat liver cell line (buffalo rat liver cells) was selected as the candidate cell line for induction of QR by a known chemopreventive agent, genistein. The choice of a liver cell line is based on the fact that liver being a major metabolic organ, phase II enzymes are commonly elaborated in liver tissues and is therefore appropriate for enzyme induction studies.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1023/A:1009706923716
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