title: Fullerene C60Penetration into Leukemic Cells and Its Photoinduced Cytotoxic Effects.
abstract: Fullerene C60as a representative of carbon nanocompounds is suggested to be promising agent for application in photodynamic therapy due to its unique physicochemical properties. The goal of this study was to estimate the accumulation of fullerene C60in leukemic cells and to investigate its phototoxic effect on parental and resistant to cisplatin leukemic cells. Stable homogeneous water colloid solution of pristine C60with average 50-nm diameter of nanoparticles was used in experiments. Fluorescent labeled C60was synthesized by covalent conjugation of C60with rhodamine B isothiocyanate. The results of confocal microscopy showed that leukemic Jurkat cells could effectively uptake fullerene C60from the medium. Light-emitting diode lamp (100 mW cm-2, λ = 420-700 nm) was used for excitation of accumulated C60. A time-dependent decrease of viability was detected when leukemic Jurkat cells were exposed to combined treatment with C60and visible light. The cytotoxic effect of photoexcited C60was comparable with that induced by H2O2, as both agents caused 50% decrease of cell viability at 24 h at concentrations about 50 μM. Using immunoblot analysis, protein phosphotyrosine levels in cells were estimated. Combined action of C60and visible light was followed by decrease of cellular proteins phosphorylation on tyrosine residues though less intensive as compared with that induced by H2O2or protein tyrosine kinase inhibitor staurosporine. All tested agents reduced phosphorylation of 55, 70, and 90 kDa proteins while total suppression of 26 kDa protein phosphorylation was specific only for photoexcited C60.The cytotoxic effect of C60in combination with visible light irradiation was demonstrated also on leukemic L1210 cells both sensitive and resistant to cisplatin. It was shown that relative value of mitochondrial membrane potential measured with tetramethylrhodamine ethyl ester perchlorate (TMRE) probe was lower in resistant cells in comparison with sensitive cells and the drop of mitochondrial potential corresponded to further decrease of resistant cell viability after C60photoexcitation. The data obtained allow to suggest that C60-mediated photodynamic treatment is a candidate for restoration of drug-resistant leukemic cell sensitivity to induction of mitochondrial way of apoptosis.
authors.affiliation: National Taras Shevchenko University of Kyiv, 64/13 Volodymyrska Street, Kyiv, 01601, Ukraine. dashaqq@gmail.com.
authors.forename: D
authors.identifier: http://orcid.org/0000-0002-1719-0512
authors.initials: D
authors.lastname: Franskevych
authors.affiliation: National Taras Shevchenko University of Kyiv, 64/13 Volodymyrska Street, Kyiv, 01601, Ukraine.
authors.forename: K
authors.initials: K
authors.lastname: Palyvoda
authors.affiliation: Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 9 Leontovicha Street, Kyiv, 01030, Ukraine.
authors.forename: D
authors.initials: D
authors.lastname: Petukhov
authors.affiliation: National Taras Shevchenko University of Kyiv, 64/13 Volodymyrska Street, Kyiv, 01601, Ukraine.
authors.forename: S
authors.initials: S
authors.lastname: Prylutska
authors.affiliation: National Taras Shevchenko University of Kyiv, 64/13 Volodymyrska Street, Kyiv, 01601, Ukraine.
authors.forename: I
authors.initials: I
authors.lastname: Grynyuk
authors.affiliation: Ilmenau University of Technology, 29 Ehrenbergstrasse, Ilmenau, 98693, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Schuetze
authors.affiliation: Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 9 Leontovicha Street, Kyiv, 01030, Ukraine.
authors.forename: L
authors.initials: L
authors.lastname: Drobot
authors.affiliation: National Taras Shevchenko University of Kyiv, 64/13 Volodymyrska Street, Kyiv, 01601, Ukraine.
authors.forename: O
authors.initials: O
authors.lastname: Matyshevska
authors.affiliation: Ilmenau University of Technology, 29 Ehrenbergstrasse, Ilmenau, 98693, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Ritter
keywords: Drug resistance; Fullerene C60; Leukemic cells; Mitochondrial membrane potential; Photodynamic therapy; Protein tyrosine phosphorylation