The polyhydroxy small-gap fullerenes [C120O30(OH)30 · 30H2O · 25Na+: SGFs] were encapsulated in multilamellar liposomes (Lpsm) composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phospho-l-serine
(DOPS), which are designated as LpsmSGFs (DOPC/DOPS/SGFs = 35 mM:15 mM:246–445 μM, diameter = 141.2 nm, ζ-potential = −35.65 mV). Radiosensitization by LpsmSGFs under X-ray irradiation was evaluated on human melanoma HMV-II cells. On 7th day after X-ray irradiation,
cell proliferation degree assessed by WST-8 decreased more markedly on cells pretreated with LpsmSGFs than Lpsm or free-SGFs. Fluorescent imaging of cells with Rhodamine123, dihydroethidium or anti-8-hydroxydeoxyguanosine antibody was monitored as an indicator for mitochondrial membrane potentials,
intracellular superoxide anion radicals (O˙−
2) or oxidative DNA-damages, respectively. After X-ray irradiation, LpsmSGFs obviously exhibited more augmented mitochondrial membrane potentials on perinuclear region of cells than Lpsm or free-SGFs. Without X-ray
irradiation, superoxide anion radicals were found principally in the cytoplasm, but, when exposed to X-ray, they were found in cell nuclei associated with oxidative DNA-damages on cells pretreated with LpsmSGFs. Meanwhile, the oxidation-reduction potentials of SGFs aqueous solution increased
by X-ray irradiation. These results suggest that LpsmSGFs-mediated generation of reactive oxygen species results in damages to cellular components such as mitochondria and DNA on cells, and thereby cell proliferation decreased. The LpsmSGFs has a potential as a pro-oxidative type radiosensitizer.
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Document Type: Research Article
Radioisotope Facilities for Medical Science, Advanced Science Research Promotion Center, Mie University, Edobashi 2-174, Tsu, Mie, 514-8507, Japan
Department of Liposome Bioengineering, Graduate School of Engineering, Mie University, Kurimamachiya-cho 1577, Tsu, Mie, 514-8507, Japan
Prefectural University of Hiroshima, Nanatsuka 562, Shobara, Hiroshima, 727-0023, Japan
Publication date: 01 June 2018
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