Introduction: 17-AAG, an Hsp90 inhibitor, exerts cytotoxic effects on several human trasformed cells and the aim of this study is to investigate the mechanism of cell death induced by 17-AAG on D22, a canine osteosarcoma (OSA) cell line, using transmission electron microscopy (TEM).Materials and Methods: D22 cell line was treated with 1, 3, 5 μM of 17-AAG at 24 and 48 hrs, fixed in 2.5% gluteraldehyde, embedded in epoxy resin and ultrathin sections, stained with uranyl acetate and lead citrate, were observed with a Zeiss EM900.Results: 17-AAG-treated cells were pleomorphic, with variable number of lamellipodia, surface bubbles and blisters, cytoplasmic vacuoles, increased RER, mitochondrial degeneration, numerous lysosomes and free ribosomes. Morphological signs of mitochondrial autophagy (mitochondria-RER complexes, isolation membranes, autophagosomes) first appeared at 24 hrs with 1 μM 17-AAG, while apoptosis was prevalent at 3 μM (24 hrs) and necrosis at 5 μM (24 hrs). Ultrastructural features of the three mechanisms of cell death appeared early after 48 hrs treatment.Conclusions: Our data demonstrate that 17-AAG exhibits a time- and dose-dependent selective cytotoxicity for OSA cells inducing different types of cell death, including the recently discovered ”mitophagy”, providing suppprt for its potential therapeutic application in clinical settings.[...]

Ultrastructural investigation of a “crime scene”: canine osteosarcoma cells killed by 17-AAG (17-allylamino-17-demethoxygeldanamycin) through autophagy/apoptosis/necrosis.

PALMIERI, CHIARA;ROMANUCCI, MARIARITA;BONGIOVANNI, LAURA;DELLA SALDA, Leonardo
2011-01-01

Abstract

Introduction: 17-AAG, an Hsp90 inhibitor, exerts cytotoxic effects on several human trasformed cells and the aim of this study is to investigate the mechanism of cell death induced by 17-AAG on D22, a canine osteosarcoma (OSA) cell line, using transmission electron microscopy (TEM).Materials and Methods: D22 cell line was treated with 1, 3, 5 μM of 17-AAG at 24 and 48 hrs, fixed in 2.5% gluteraldehyde, embedded in epoxy resin and ultrathin sections, stained with uranyl acetate and lead citrate, were observed with a Zeiss EM900.Results: 17-AAG-treated cells were pleomorphic, with variable number of lamellipodia, surface bubbles and blisters, cytoplasmic vacuoles, increased RER, mitochondrial degeneration, numerous lysosomes and free ribosomes. Morphological signs of mitochondrial autophagy (mitochondria-RER complexes, isolation membranes, autophagosomes) first appeared at 24 hrs with 1 μM 17-AAG, while apoptosis was prevalent at 3 μM (24 hrs) and necrosis at 5 μM (24 hrs). Ultrastructural features of the three mechanisms of cell death appeared early after 48 hrs treatment.Conclusions: Our data demonstrate that 17-AAG exhibits a time- and dose-dependent selective cytotoxicity for OSA cells inducing different types of cell death, including the recently discovered ”mitophagy”, providing suppprt for its potential therapeutic application in clinical settings.[...]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11575/16890
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