INTRODUCTION In this work we investigated the biological activity of a new synthesized series of guanylhydrazone derivatives. The compounds under test bear the guanidine group as other drugs (i.e. mitoguazone or methyl-glyoxal-bisguanylhydrazone), which have been usefully employed in clinical trials in the treatment of malignant lymphoma, head, neck and esophageal cancer. Mitoguazone is known to induce apoptosis in different human cancer cell lines in a concentration and time dependent way. Previous studies have shown that the inhibition of mitochondrial functions is a feature of many guanidino-containing drugs[1]. Therefore, we investigated the cytostatic and cytotoxic effects of seven new derivatives in HL60 cells, and in particular if disruption of the Dym was involved in their antiproliferative action. METHODS The cells were treated at 2x105cell/mL with the compounds under test dissolved in DMSO. After 24hr of treatment cells growth was monitored and cell cycle distribution was evaluated by flow cytometry. Cell death was characterized by monitoring spectrophotometrically caspase activation, ATP depletion and LDH release. Dym, ROS production and cell viability were detected by flow cytometry. RESULTS 2mM treatment of cells with all tested compounds had only cytostatic effects, without significant variation on cell viability, ROS production, Dym and on cell cycle cell distribution. The study was then focused on the two more active molecules, HD and 36. At concentration higher than 2mM these compounds induced cell death, characterized by apoptotic features as chromatin condensation and effector caspases activation. Furthermore loss of Dym and ROS production were yet detectable after 1hr of treatments: the latter effect was more remarkable for compound 36, which was able to increase both peroxide and superoxide species, than for HD which was able to increase only peroxide production. Finally, the cytotoxic effect was confirmed by the ATP depletion and by LDH release in the extracellular milieu. In conclusion the collapse of Dym accompanied by a decrease of cellular ATP content indicates an interference of these molecules with cellular energetics. Further studies are ongoing to clarify the exact mechanism for the observed cytotoxic effects. [1] Syntesis and antitumor activity od guanylhydrazones from 6-(2,4-Dichloro-5-nitrophenyl)imidazo[2,1-b]thiazoles and 6-Pyridylimidazo[2,1-b]thiazoles. Andreani et al. J.Med.Chem. 2006, 49, 7897.

CYTOSTATIC AND CYTOTOXIC EFFECTS OF A NEW SYNTHESIZED GUANYLHYDRAZONE DERIVATIVES IN HUMAN LEUKEMIC CELL LINE HL-60

FARRUGGIA, GIOVANNA;CAPPADONE, CONCETTINA;CALONGHI, NATALIA;DI GIORGIO, FRANCESCA;MANGANO, CHIARA;PAROLIN, CAROLA ELEONORA;MASOTTI, LANFRANCO
2007

Abstract

INTRODUCTION In this work we investigated the biological activity of a new synthesized series of guanylhydrazone derivatives. The compounds under test bear the guanidine group as other drugs (i.e. mitoguazone or methyl-glyoxal-bisguanylhydrazone), which have been usefully employed in clinical trials in the treatment of malignant lymphoma, head, neck and esophageal cancer. Mitoguazone is known to induce apoptosis in different human cancer cell lines in a concentration and time dependent way. Previous studies have shown that the inhibition of mitochondrial functions is a feature of many guanidino-containing drugs[1]. Therefore, we investigated the cytostatic and cytotoxic effects of seven new derivatives in HL60 cells, and in particular if disruption of the Dym was involved in their antiproliferative action. METHODS The cells were treated at 2x105cell/mL with the compounds under test dissolved in DMSO. After 24hr of treatment cells growth was monitored and cell cycle distribution was evaluated by flow cytometry. Cell death was characterized by monitoring spectrophotometrically caspase activation, ATP depletion and LDH release. Dym, ROS production and cell viability were detected by flow cytometry. RESULTS 2mM treatment of cells with all tested compounds had only cytostatic effects, without significant variation on cell viability, ROS production, Dym and on cell cycle cell distribution. The study was then focused on the two more active molecules, HD and 36. At concentration higher than 2mM these compounds induced cell death, characterized by apoptotic features as chromatin condensation and effector caspases activation. Furthermore loss of Dym and ROS production were yet detectable after 1hr of treatments: the latter effect was more remarkable for compound 36, which was able to increase both peroxide and superoxide species, than for HD which was able to increase only peroxide production. Finally, the cytotoxic effect was confirmed by the ATP depletion and by LDH release in the extracellular milieu. In conclusion the collapse of Dym accompanied by a decrease of cellular ATP content indicates an interference of these molecules with cellular energetics. Further studies are ongoing to clarify the exact mechanism for the observed cytotoxic effects. [1] Syntesis and antitumor activity od guanylhydrazones from 6-(2,4-Dichloro-5-nitrophenyl)imidazo[2,1-b]thiazoles and 6-Pyridylimidazo[2,1-b]thiazoles. Andreani et al. J.Med.Chem. 2006, 49, 7897.
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Farruggia G.; Cappadone C.; Calonghi N.; Di Giorgio F.; Mangano C.; Parolin C.; Masotti L.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/57105
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