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For selection and identification of potential chemopreventive agents, a biochemical assay using induction of a phase II enzyme, glutathione-S-transferase (GST) in a liver cell culture is described. A normal human liver cell line (Chang liver cells) was selected as the candidate cell line for induction of GST (liver tissues are abundant in GST) by a known chemopreventive agent, oltipraz. Exponentially growing cells plated for 24 hours are exposed to various doses of a chemopreventive agent for an additional 24 hours, homogenized by sonication, and the homogenate is assayed for GST in a modified microplate enzyme assay using CDNB (1-chloro-2,4-dinitrobenzene) as a substrate. A concurrent protein assay is performed to determine the specific enzyme activity. As the assay is modified from a spectrophotometric assay to a microplate assay, it is reliable, sensitive and fast, a significant number of test agents can be screened in a short time. |
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