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© 1993 Oxford University Press

research-article

Validation of two test systems for detecting tumor promoters and EBV inducers: comparative responses of several agents in DR-CAT Raji cells and in human granulocytes

Guy Bouvier, Manfred Hergenhahn 2, Axel Polack 1, Georg W. Bornkamm 1 and helmut Bartsch 3

International Agency For Research on Cancer 150 Cours Albert Thomas, 69372 Lyon Cedex 08
1GSF Forschungszentrum Hamatologikum Marchioninistraße 25, D-8000 Mûnchen 70, Germany
2Visiting scientist on leave from the German Cancer Research Center D-69 Heidelberg, Germany

3To whom all correspondence should be addressed

The response to a number of agents has been compared in two short-term assays used for the detection of virus inducers and tumor promoters: (i) induction of the EBV-DR-promoter in Raji cells, as measured by DR-CAT induction (DR-CAT test) and (ii) induction of the oxidative burst in human PMN, as measured by chemiluminescence in the presence of luminol or lucigenin (CL test). In order to validate the two assays, we have investigated the responses to 12-O-tetradecanoyl-phorbol-13-acetate (TPA), 1, 2-dioctanoylglycerol (DAG), phospholipase C (PLC EC-3–1-4–30) and ionophore A23187 [GenBank] , which are active in both systems; arachidonic acid, linoleic acid and NaCl were found active only in the CL test. Staurosporine (protein kinase inhibitor), tamoxifen (estrogen antagonist and protein kinase C inhibitor), forskolin (protein kinase A activator), R59949 [GenBank] (diacylglycerol kinase inhibitor), curcumin and N-acetyl-L-cysteine (scavengers of reactive oxygen species) and NaCl acted as inhibitors. A good concordance of the EC50 values of inducing substances was found between the two assays, except for A23187 [GenBank] and DAG, which were required at much higher concentrations in the DR-CAT test. The inhibition patterns by the panel of inhibitors revealed similarities and discrepancies in the induction pathways between the two systems, providing information on their mode of action. The two assays, which complement each other, were shown to detect a number of known or suspected EBV inducers or tumor promoters, and thus appear useful for screening of new compounds or mixtures as well as of potential antiviral and antipromoting substances.


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