Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene

被引:227
作者
Isakovic, A
Markovic, Z
Todorovic-Markovic, B
Nikolic, N
Vranjes-Djuric, S
Mirkovic, M
Dramicanin, M
Harhaji, L
Raicevic, N
Nikolic, Z
Trajkovic, V
机构
[1] Univ Belgrade, Sch Med, Inst Microbiol & Immunol, YU-11000 Belgrade, Serbia
[2] Univ Belgrade, Sch Med, Inst Biochem, YU-11000 Belgrade, Serbia
[3] Vinca Inst Nucl Sci, YU-11000 Belgrade, Serbia
[4] Inst Biol Res, YU-11000 Belgrade, Serbia
[5] Univ Belgrade, Fac Phys, YU-11000 Belgrade, Serbia
关键词
cytotoxicity; fullerene; C60 reactive oxygen species; apoptosis; necrosis;
D O I
10.1093/toxsci/kfj127
中图分类号
R99 [毒物学(毒理学)];
学科分类号
100405 ;
摘要
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.
引用
收藏
页码:173 / 183
页数:11
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