In the research effects of bergapten (5-MOP) and peucedanin, furanocoumarins isolated from fruits of Peucedanum tauricum Bieb. (Apiaceae), on apoptosis, necrosis and heat-shock proteins Hsp 27 and Hsp 72 expression in human carcinoma cells HeLa B (ECACC No 85060701) were preliminary studied. The purity of analysed compounds was confirmed by RP HPLC, EI-MS and 1D, 2D NMR. For apoptosis and necrosis detection the mixture of Hoechst 33342 and propidium iodide (PI) was used. Early apoptosis was detected using Annexin V Test. Cells were incubated with furanocoumarins at the concentrations of 1, 5, 10 and 15 μg/ml. The level of heat-shock protein expression was determined by Western blot technique. Quantitative analysis of Hsp 27 and Hsp 72 was done with the Bio-Profil Bio-1D Windows Application V.99.03 program. The obtained results were analysed for significance by one-way ANOVA test. As a result of our experiments we found that peucedanin was more effective in apoptosis induction than bergapten. In HeLa cells treated with peucedanin only low necrosis was observed. The incubation with 15 μg/ml of peucedanin for 24 hours inhibited Hsp 72 expression in cells by 77.5% (± 0.05), and Hsp 27 by 74.0% (± 0.02) after five hours of treatment as compared to the control. The inhibition of Hsps expression after peucedanin treatment may be responsible for increased apoptosis induction (11 .4%) detected by Annexine V test. Incubation with furanocoumarins has an influence on morphology of HeLa cells (many cytoplasmic protrudings and bulges of “blebbing” cytoplasm were observed).
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