ANTIPROLIFERATIVE EFFECT OF POLYAMINES AND PUTRESCINE AND THEIR USE AS BIOCHEMICAL MARKERS OF THE BIOSYNTHESIS RATE IN L-CELL CULTURE

Effects of inhibitors of enzymes involved in biosynthesis of putrescine and polyamines on growth of mice C3H strain transformed fibroblasts as well as on the intracellular content of putrescine and polyamines were studied. Putrescine and polyamines were estimated as dansyl derivatives using HPLC. DL-alpha-difluoromethyl ornithine (F2MeOrn) and methyl glyoxalbis (quanyl hydrazone) (MGBG) were added to cultivation medium at final concentration 10(-4) M daily within 4 days. The cell culture was not treated within the following 3 days of the experiment. Distinct exceeding of spermidine content over concentrations of putrescine and spermine proved to be the specific property of L-cells growth in the culture. Amount of cells correlated strongly with intracellular content of spermidine and spermine (r = 0.964). Treatment of L-cells with F2MeOrn singly or in combination with MGBG led to 4- and 6-fold decrease in their growth rate, respectively. F2MeOrn altered content of polycations 2 days earlier in the cells which exhibited distinct correlation between putrescine and polyamines (r = 0.943). The rate of MGBG inhibition correlated distinctly with content of putrescine and polyamines (r = 0.953) as well as strong correlation between putrescine and polyamines content (r = 0.994) was detected after simultaneous effect of these inhibitors. This suggests that polyamines were involved in the antiproliferative action of the inhibitors studied and may be used as markers of cytostatics chemotherapeutic efficiency.