CELLULAR & MOLECULAR BIOLOGY LETTERS Volume 6, (2001) pp 649 Ą 675 Received 23 July 2001 Accepted 10 September 2001 * Corresponding author, fax: (+4871) 3479211,e-mail: kaz@basmed.am.wroc.pl IMPACT OF FOUR ANTIMUTAGENS ON APOPTOSIS IN GENOTOXICALLY DAMAGED LYMPHOCYTES IN VITRO KAZIMIERZ GASIOROWSKI 1* , BARBARA BROKOS 1 , ANNA KULMA 2 , ANTONI OGORZAÓEK 3 and KATARZYNA SKORKOWSKA 1 1 Wroc’aw Medical University, Department of Basic Medical Sciences, Kochanowskiego 14, 51- 601 Wroc’aw, Poland, 2 University of Wroc’aw, Department of Genetic Biochemistry, Przybyszewskiego 63/77, 51-148 Wroc’aw, Poland, 3 University of Wroc’aw, Department of Zoology, Sienkiewicza 21, 50-335 Wroc’aw, Poland Abstract: An antimutagenic activity of fluphenazine, todralazine, anthocyanins and alkylresorcinols was established in a battery of short-term cytogenetic tests. One of the possible mechanisms of their antimutagenic action could be an increase in apoptotic elimination of heavily-damaged cells from a culture. In this paper we provide data on quantitative estimation of the antimutagensµ impact on apoptosis in lymphocyte cultures exposed in the G 0 -phase to genotoxic agents: hydrogen peroxide (0.2mM, 20 min.) or benzo[a]pyrene (40 óM, 90 min.), and then cultured for 36 hrs in the presence of a lectin (PHA-M, 1% v/v) and each of the tested antimutagens. Apoptosis was estimated by means of microscopic examination of cell smears stained with a mixture of fluorochromes (ethidium bromide/acridine orange) as well as of the results of DNA separation with the field inversion gel electrophoresis. By microscopic examination we assessed that the frequencies of cells exhibiting morphological features of apoptosis considerably increased in the cultures containing the antimutagens. The FIGE separation of DNA from those cultures proved that the DNA content in the 30-50 kb domain was markedly elevated, as compared with the control cultures that did not contain antimutagens. It was established in the regression analysis that the apoptosis-enhancing effect significantly depended on the concentration of each tested antimutagen in a culture medium. However, marked differences of apoptosis-enhancig potency were noticed among the four antimutagens. The multicriterial analysis proved that the apoptosis-enhancing