engleski

Molecular alterations induced with diazene JK-279 in human cervical carcinoma cells

Glutathione (GSH) plays an essential role in cellular homeostasis (1,2). It protects cells against free radical induced oxydant injury and detoxificates numerous endogenous and foreign electrophilic compounds. New compounds diazenecarboxamides (diazenes) have been synthesized that should, according to their structure and biochemical properties, lower the intracellular GSH content (3), thus inhibiting the growth of tumor cells. We have shown previously (4), that from ten new synthesized diazenecarboxamides (diazenes), diazene JK-279 was mostly effective. It reduced the cell survival of all ten cell lines examined: cervical carcinoma, laryngeal carcinoma, mammary carcinoma, breast adenocarcinoma and glioblastoma cells, as well as their drug-resistant cells sublines. The aim of the present study was to elucidate the molecular mechanisms involved in this phenomenon. Human cervical carcinoma HeLa cells were used as the experimental system. The kinetic of apoptosis induction was followed after the treatment with JK-279 by flourescent microscope for 72 h posttreatment period (5). The occurrence of apoptosis was confirmed by analysing DNA fragmentation in agarose gel (6). For the cell cycle phase determination, flow cytometric analysis was performed by the standard procedure, using propidium iodide for staining and EPICS-computer PARA1 program for analysis of the data. Results show that diazene JK-279 induced significant degradation of the genomic DNA into internucleosomal sized DNA fragments (even one hour after the treatment), indicating apoptosis. The treatment with diazene JK-279 resulted in accumulation of cells in S and G2-M phases, that was mostly expressed 12 h after the treatment. Our results show that diazene JK-279 arrested the cells in S and G2 phase of the cell cycle, and induced apoptosis in human cervical carcinoma cells. (1) A. Meister, Cancer Res., 54, 1969-1975 (1994) (2) M.F.C.M. Knapen, P.L.M. Zusterzeel, W.H.M. Peters, E.A.P. Stegeers, Eur.J.Obstet.Gynecol. Reprod.Biol., 82, 171-184 (1999) (3) J. Košmrlj, M. Kočevar, S. Polanc, J.Chem.Soc.Perkin.Trans., 1, 3917-3919 (1998) (4) M. Osmak, T. Bordukalo, B. Jernej, J. Košmrlj, S. Polanc, Anti-Cancer Drugs, 10, 853-859 (1999) (5) M. Osmak, M. Abramić, A. Brozović, M. Hadžija, Period.biol. 99(3), 329-333 (1997) (6) M.F. Ausubel, R. Brent, R.E. Kingston, D.D. More, J.G. Seidman, J.A. Smith, K. Struhl, Short Protocols in Molecular Biology, 2nd ed.

diazene; tumor cells; apoptosis

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