Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene
Authors
Isaković, Aleksandra J.Marković, Zoran M.
Todorović-Marković, Biljana
Nikolić, Nadežda S.
Vranješ-Đurić, Sanja
Mirković, Marija D.
Dramićanin, Miroslav
Harhaji, Ljubica M.
Raičević, Nevena
Nikolić, Zoran M.
Trajković, Vladimir S.
Article (Published version)
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The mechanisms underlying the cytotoxic action of pure fullerene suspension (nano-C-60) and water-soluble polyhydroxylated fullerene [C-60(OH)(n)] were investigated. Crystal violet assay for cell viability demonstrated that nano-C-60 was at least three orders of magnitude more toxic than C-60(OH)(n) to mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines. Flow cytometry analysis of cells stained with propidium iodide (PI), PI/annexin V-fluorescein isothiocyanate, or the redox-sensitive dye dihydrorhodamine revealed that nano-C-60 caused rapid (observable after few hours), reactive oxygen species (ROS)-associated necrosis characterized by cell membrane damage without DNA fragmentation. In contrast, C-60(OH)(n) caused delayed, ROS-independent cell death with characteristics of apoptosis, including DNA fragmentation and loss of cell membrane asymmetry in the absence of increased permeability. Accordingly, the antioxidant N-acetylcysteine protected the cell lines from n...ano-C-60 toxicity, but not C-60(OH)(n) toxicity, while the pan-caspase inhibitor z-VAD-fmk blocked C-60(OH)(n)-induced apoptosis, but not nano-C-60-mediated necrosis. Finally, C-60(OH)(n) antagonized, while nano-C-60 synergized with, the cytotoxic action of oxidative stress-inducing agents hydrogen peroxide and peroxynitrite donor 3-morpholinosydnonimine. Therefore, unlike polyhydroxylated C-60 that exerts mainly antioxidant/cytoprotective and only mild ROS-independent pro-apoptotic activity, pure crystalline C-60 seems to be endowed with strong pro-oxidant capacity responsible for the rapid necrotic cell death.
Keywords:
cytotoxicity / fullerene / C60 reactive oxygen species / apoptosis / necrosisSource:
Toxicological Sciences, 2006, 91, 1, 173-183Funding / projects:
DOI: 10.1093/toxsci/kfj127
ISSN: 1096-6080
PubMed: 16476688
WoS: 000236808200020
Scopus: 2-s2.0-33645806308
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VinčaTY - JOUR AU - Isaković, Aleksandra J. AU - Marković, Zoran M. AU - Todorović-Marković, Biljana AU - Nikolić, Nadežda S. AU - Vranješ-Đurić, Sanja AU - Mirković, Marija D. AU - Dramićanin, Miroslav AU - Harhaji, Ljubica M. AU - Raičević, Nevena AU - Nikolić, Zoran M. AU - Trajković, Vladimir S. PY - 2006 UR - https://vinar.vin.bg.ac.rs/handle/123456789/3002 AB - The mechanisms underlying the cytotoxic action of pure fullerene suspension (nano-C-60) and water-soluble polyhydroxylated fullerene [C-60(OH)(n)] were investigated. Crystal violet assay for cell viability demonstrated that nano-C-60 was at least three orders of magnitude more toxic than C-60(OH)(n) to mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines. Flow cytometry analysis of cells stained with propidium iodide (PI), PI/annexin V-fluorescein isothiocyanate, or the redox-sensitive dye dihydrorhodamine revealed that nano-C-60 caused rapid (observable after few hours), reactive oxygen species (ROS)-associated necrosis characterized by cell membrane damage without DNA fragmentation. In contrast, C-60(OH)(n) caused delayed, ROS-independent cell death with characteristics of apoptosis, including DNA fragmentation and loss of cell membrane asymmetry in the absence of increased permeability. Accordingly, the antioxidant N-acetylcysteine protected the cell lines from nano-C-60 toxicity, but not C-60(OH)(n) toxicity, while the pan-caspase inhibitor z-VAD-fmk blocked C-60(OH)(n)-induced apoptosis, but not nano-C-60-mediated necrosis. Finally, C-60(OH)(n) antagonized, while nano-C-60 synergized with, the cytotoxic action of oxidative stress-inducing agents hydrogen peroxide and peroxynitrite donor 3-morpholinosydnonimine. Therefore, unlike polyhydroxylated C-60 that exerts mainly antioxidant/cytoprotective and only mild ROS-independent pro-apoptotic activity, pure crystalline C-60 seems to be endowed with strong pro-oxidant capacity responsible for the rapid necrotic cell death. T2 - Toxicological Sciences T1 - Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene VL - 91 IS - 1 SP - 173 EP - 183 DO - 10.1093/toxsci/kfj127 ER -
@article{ author = "Isaković, Aleksandra J. and Marković, Zoran M. and Todorović-Marković, Biljana and Nikolić, Nadežda S. and Vranješ-Đurić, Sanja and Mirković, Marija D. and Dramićanin, Miroslav and Harhaji, Ljubica M. and Raičević, Nevena and Nikolić, Zoran M. and Trajković, Vladimir S.", year = "2006", abstract = "The mechanisms underlying the cytotoxic action of pure fullerene suspension (nano-C-60) and water-soluble polyhydroxylated fullerene [C-60(OH)(n)] were investigated. Crystal violet assay for cell viability demonstrated that nano-C-60 was at least three orders of magnitude more toxic than C-60(OH)(n) to mouse L929 fibrosarcoma, rat C6 glioma, and U251 human glioma cell lines. Flow cytometry analysis of cells stained with propidium iodide (PI), PI/annexin V-fluorescein isothiocyanate, or the redox-sensitive dye dihydrorhodamine revealed that nano-C-60 caused rapid (observable after few hours), reactive oxygen species (ROS)-associated necrosis characterized by cell membrane damage without DNA fragmentation. In contrast, C-60(OH)(n) caused delayed, ROS-independent cell death with characteristics of apoptosis, including DNA fragmentation and loss of cell membrane asymmetry in the absence of increased permeability. Accordingly, the antioxidant N-acetylcysteine protected the cell lines from nano-C-60 toxicity, but not C-60(OH)(n) toxicity, while the pan-caspase inhibitor z-VAD-fmk blocked C-60(OH)(n)-induced apoptosis, but not nano-C-60-mediated necrosis. Finally, C-60(OH)(n) antagonized, while nano-C-60 synergized with, the cytotoxic action of oxidative stress-inducing agents hydrogen peroxide and peroxynitrite donor 3-morpholinosydnonimine. Therefore, unlike polyhydroxylated C-60 that exerts mainly antioxidant/cytoprotective and only mild ROS-independent pro-apoptotic activity, pure crystalline C-60 seems to be endowed with strong pro-oxidant capacity responsible for the rapid necrotic cell death.", journal = "Toxicological Sciences", title = "Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene", volume = "91", number = "1", pages = "173-183", doi = "10.1093/toxsci/kfj127" }
Isaković, A. J., Marković, Z. M., Todorović-Marković, B., Nikolić, N. S., Vranješ-Đurić, S., Mirković, M. D., Dramićanin, M., Harhaji, L. M., Raičević, N., Nikolić, Z. M.,& Trajković, V. S.. (2006). Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene. in Toxicological Sciences, 91(1), 173-183. https://doi.org/10.1093/toxsci/kfj127
Isaković AJ, Marković ZM, Todorović-Marković B, Nikolić NS, Vranješ-Đurić S, Mirković MD, Dramićanin M, Harhaji LM, Raičević N, Nikolić ZM, Trajković VS. Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene. in Toxicological Sciences. 2006;91(1):173-183. doi:10.1093/toxsci/kfj127 .
Isaković, Aleksandra J., Marković, Zoran M., Todorović-Marković, Biljana, Nikolić, Nadežda S., Vranješ-Đurić, Sanja, Mirković, Marija D., Dramićanin, Miroslav, Harhaji, Ljubica M., Raičević, Nevena, Nikolić, Zoran M., Trajković, Vladimir S., "Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene" in Toxicological Sciences, 91, no. 1 (2006):173-183, https://doi.org/10.1093/toxsci/kfj127 . .