Tag Archives: fullerene C(60)

Activation of store – operated Ca(2+) entry in cisplatin resistant leukemic cells after treatment with photoexcited fullerene C(60) and cisplatin

D. V. Franskevych, I. I. Grynyuk, S. V. Prylutska, O. P. Matyshevska

Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: dashaqq@gmail.com

Ca2+-regulating system in cancer cells is suggested to be remodulated particularly by reduced store-operated Ca2+ entry (SOCE) through plasma membrane in order to maintain moderately reduced cytosolic Ca2+ concentration and to avoid apoptosis. The endoplasmic reticulum (ER) Ca2+ pool content and the size of SOCE in leukemic wild type (L1210) and resistant to cisplatin (L1210R) cells in control, after treatment with either cisplatin (1 µg/ml) or photoexcited fulleren C60 (10-5 M) alone, or their combination were estimated with the use of Indo-1 AM. The SOCE in resistant to cisplatin L1210R cells was found to be lower than in the wild-type cells. After treatment with cisplatin the decrease of thapsigargin (TG)-sensitive ER Ca2+ pool with no significant increase of SOCE was observed in L1210 cells, while no changes were detected in L1210R cells. Photoexcitation of intracellular accumulated fullerene C60 in the visible range of spectrum (410-700 nm) was accompanied by increase of SOCE not only in sensitive, but in resistant cells as well. In resistant L1210R cells treated with photoexcited C60 essential effect of cisplatin on Ca2+ homeostasis became obvious: the size of SOCE proved to be higher than after treatment with photoexcited C60 alone. The data obtained allow suggesting­ the influence of photoexcited C60 not only on Ca2+-regulating system, but on those involved in controlling cisplatin entry into drug resistant cancer cells.

Photoactivated fullerene C(60) induces store-operated Ca(2+) entry and cytochrome c release in Jurkat cells

S. M. Grebinyk1, K. O. Palyvoda2, S. V. Prylutska1, I. I. Grynyuk1,
A. A. Samoylenko2, L. B. Drobot2, O. P. Matyshevska1

1Taras Shevchenko Kyiv National University, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: matysh@yahoo.com

The values of endoplasmic reticulum Ca2+-pool and store-operated Ca2+ entry (SOCE) were estimated in rat thymocytes and Jurkat cells loaded with indo-1 and treated with thapsigargin. It was shown that the relative value of SOCE in thymocytes was substantially lower than in Jurkat cells. Significant increase of SOCE in Jurkat cells preincubated with 10-5 M C60 and exposed to uv/visible light irradiation was detected at 1-3 h after exposure. At this time FCCP-induced Ca2+-release from mitochondria was shown to be reduced, while cytochrome c level into the cytoplasm of Jurkat cells, detected by Western blot analysis, to be increased. It is supposed that Ca2+ flux remodulation induced by photoexcited fullerene C60 in Jurkat cells might be involved in the initiation of signalling events leading to cell apoptosis.

Generation of active oxygen forms in rat tymocytes under action of hydrogen peroxide and fullerene C(60)

S. M. Grebinyk, I. I. Grynyuk, S. V. Prylutska, O. P. Matyshevska

Taras Shevchenko Kyiv National University, Ukraine;
e-mail: grebnik_z@yahoo.com

The dynamics of active oxygen forms (AOF) generation in rat thymocytes 50 min after treatment with 0.1 and 0.5 mM H2O2 was estimated with the use of fluorescent probe DCFDA. Both enhanced AOF generation, which was dependent on H2O2 concentration, and glutathione peroxidase and superoxide dismutase activation, followed by a decrease of thymocytes viability were demonstrated.
Preincubation of cells with 10-5 M fullerene C60 was shown not only to prevent H2O2 – induced AOF generation but to increase viability of H2O2-treated thymocytes at more prolonged time period. The data obtained indicate to fullerene C60 ability to prevent oxidative stress in thymocytes.

Modulation of cisplatin-induced reactive oxygen species production by fullerene C(60) in normal and transformed lymphoid cells

D. V. Franskevych, I. I. Grynyuk, S. V. Prylutska, O. P. Matyshevska

Taras Shevchenko National University of Kyiv, Ukraine;
е-mail: dashaqq@gmail.com

The early response of normal (Wistar rat thymocytes) and transformed (mice lymphoid leukemia L1210) cells to treatment with anticancer drug cisplatin or to combined treatment with cisplatin and carbon nanostructure fullerene C60 was studied. We demonstrated with fluorescent probes DCFH-DA and TMRE that cisplatin at concentration 1 μg/ml induced reactive oxygen species (ROS) production and decreased the value of mitochondrial membrane potential in both cell types. The combined treatment with cisplatin (1 μg/ml) and fullerene C60 (7.2 μg/ml) was shown to be followed by oppositely directed modulation of ROS production in thymocytes and L1210 cells. Cisplatin-induced ROS production was intensified in L1210 cells, while in thymocytes it was decreased. It is supposed that the different effects of combined treatment are associated with peculiarities of fullerene C60 accumulation and localization in normal and cancer cells.

Effect of fullerene C(60) on ATPase activity and superprecipitation of skeletal muscle actomyosin

K. S. Andreichenko1, S. V. Prylutska1, K. O. Medynska1, K. I. Bogutska1,
N. E. Nurishchenko1, Yu. I. Prylutskyy1, U. Ritter2, P. Scharff2

Joint Ukrainian-German Center on Nanobiotechnology
1Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: prylut@ukr.net;
2Technical University of Ilmenau, Institute of Chemistry
and Biotechnology, Germany

Creation of new biocompatible nanomaterials, which can exhibit the specific biological effects, is an important complex problem that requires the use of last accomplishments of biotechnology. The effect of pristine water-soluble fullerene C60 on ATPase activity and superprecipitation reaction of rabbit skeletal muscle natural actomyosin has been revealed, namely an increase of actomyosin superprecipitation and Мg2+, Са2+– and K+-ATPase activity by fullerene was investigated. We conclude that this finding offers a real possibility for the regulation of contraction-relaxation of skeletal muscle with fullerene C60.