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C60 fullerene enhances cisplatin anticancer activity and overcomes tumor cell drug resistance

Nano Research 2017年第2期10卷 652-671页

作者： Svitlana Prylutska rostyslav Panchuk Grzegorz Golunski Larysa Skivka Yuriy Prylutskyy Vasyl Hurmach Nadya Skorohyd Agnieszka Borowik Anna Woziwodzka Jacek Piosik Olena Kyzyma Vasil Garamus Leonid Bulavin Maxim Evstigneev Anatoly Buchelnikov rostyslav stoika Walter Berger Uwe Ritter Peter Scharff Taras Shevchenko National University of Kyiv 64 Volodymyrska Str. 01601 Kyiv Ukraine Institute of Cell Biology NAS of Ukraine 14-16 Drahomanov 5tr. 79005 Lviv Ukraine Laboratory of Biophysics Intercollegiate Faculty ofBiotechnology UG-MUG Abrahama 58 80-307 Gdansk Poland Joint Institute for Nuclear Research 6 Joliot-Curie Str. 141980 Dubna Moscow reg. Russia Helmholtz-Zentrum Geesthacht: Centre for Materials and Coastal Research 1 Max-Planck Str. 21502 Geesthacht Germany Belgorod State University 85 Pobedy Str. 308015 Belgorod Russia Institute of Cancer Research and Comprehensive Cancer Center Medical University Vienna 1090 8A Borschkegasse Vienna Austria Institute of Chemistry and Biotechnology Technical University ofllmenau 25 Weimarer Str. 98693 Ilmenau Germany

We formulated and analyzed a novel nanoformulation of the anticancer drug cisplatin （Cis） with C60 fullerene （C60＋Cis complex） and showed its higher toxicity toward tumor cell lines in vitro when compared to Cis alone. The highest toxicity of the complex was observed in HL-60/adr and HL-60/vinc chemotherapy- resistant human leukemia cell sublines （resistant to Adriamycin and Vinculin, respectively）. We discovered that the action of the C60＋Cis complex is associated with overcoming the drug resistance of the tumor cell lines through observing an increased number of apoptotic cells in the Annexin V/PI assay. Moreover, in vivo assays with Lewis lung carcinoma （LLC） C57BL/6J male mice showed that the C60＋Cis complex increases tumor growth inhibition, when compared to Cis or C60 fullerenes alone. Simultaneously, we conducted a molecular docking study and performed an Ames test. Molecular docking specifies the capability of a C60 fullerene to form van der Waals interactions with potential binding sites on P-glycoprotein （P-gp）, multidrug resistance protein 1 （MRP-1）, and multidrug resistance protein 2 （MRP-2） molecules. The observed phenomenon revealed a possible mechanism to bypass tumor cell drug resistance by the C60＋Cis complex. Additionally, the results of the Ames test show that the formation of such a complex diminishes the Cis mutagenic activity and may reduce the probability of secondary neoplasm formation. In conclusion, the C60＋Cis complex effectively induced tumor cell death in vitro and inhibited tumor growth in vivo, overcoming drug resistance likely by the potential of the C60 fullerene to interact with P-gp, MRP-1, and MRP-2 molecules. Thus, the C60＋Cis complex might be a potential novel chemotherapy modification.

关键词： molecular docking,small-angle X-ray scattering,apoptosis,mutagenic activity lung carcinoma (LLC),cytotoxicity

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Histone H1/MBP hydrolysing antibodies - novel potential marker in diagnosis of disease severity in systematic lupus erythematosus patients

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Health 2010年第10期2卷 1204-1207页

作者： Valentyna Chopyak Yaroslav Tolstiak Iryna Magoryvska rostyslav Bilyy rostyslav Bilyy Natalya Korniy Yuriy Kit rostyslav stoika 不详

Recently we have shown the presence of catalytically active IgGs, capable to cleave histone H1 and bovine myelin basic protein (MBP), in blood serum of SLE patients. Here we present data that demonstrate the correlation between a) proteolytic activity towards histone H1 and MBP of IgG-antibodies from blood serum of SLE patients and b) disease severity level in these patients. IgGs were isolated from blood serum by chromatography on protein G-sepharose. Commercial preparations of bovine myelin basic proteins (MBP) and calf thymus histone H1 were used as substrates. Analysis of the proteolytic activity showed that 16 of 38 lgG-preparations (42,1%) obtained from blood serum of SLE patients were capable of cleaving both histone H1 and MBP with different efficiency. It was revealed that the presence in blood serum of lgGs possessing proteolytic activity towards both histone H1 and bMBP closely correlates with manifestation of the disease severity in SLE patients.

关键词： System Lupus Erythematosus Proteolytic Activity IgG-Antibodies Disease Severity

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Nemeth-Kellner Lymphoma Is a Valid Experimental Model in Testing Chemical Agents for Anti-Lymphoproliferative Activity

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Open Journal of Blood Diseases 2013年第3期3卷 1-6页

作者： Maxim D. Lootsik Maksym M. Lutsyk rostyslav S. stoika Department of Histology Cytology and Embryology Danylo Halytsky National Medical University Lviv Ukraine. Institute of Cell Biology National Academy of Sciences of Ukraine Lviv Ukraine nstitute of Cell Biology National Academy of Sciences of Ukraine Lviv Ukraine

Scheme of detection and investigation of antitumor activity of chemical substances by use of experimental murine Nemeth-Kellner lymphoma (NK/Ly) are described. It includes monitoring of growth of NK/Ly ascites by everyday weighting of animals in course of intraperitoneal application of tested substance in 24 or 48 hours after inoculation of 107 tumor cells. The inhibitory effect became distinct on the 6th-8th day of experiment, and 3 types of response were observed: 1) complete suppression of ascites development, corresponding to high antitumor activity of tested substance;2) partial suppression of ascites growth (weak antitumor activity);3) insignificant effect on ascites growth (lack of activity, at least towards hematoblastoses). As compared with leukemia L1210 or P388 lymphoma NK/Ly provides more distinct daily increment of body weight due to tumor growth. Clinical pattern of tumor development was supported by cytomorphological investigations of tumor cells which included: 1) cells count in ascites;2) measuring cell dimensions and ratio of enlarged cells in a population (diameter over 17 μm in case of NK/Ly);3) quantitative evaluation of tumor cells damage by smears count after staining with azure-eosin, bromophenol blue and hematoxylin. In case of complete suppression of ascites growth the cytological investigation was conducted by delayed application of tested agent to animals with the developed ascites. After treatment with vinblastine or doxorubicin NK/Ly cells represent a convenient object for fractionation of cell population, as demonstrated by separation of giant and small tumor cell subpopulations.

关键词： Nemeth-Kellner Lymphoma Antitumor Substances Screening

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