Nanochemistry and biotechnology
Antitumor activity of C60 fullerene-Cisplatin nanocomplex against Lewis lung carcinoma cells
Prylutska S.V.1, Grynyuk I.I.1, Skaterna T.D.2, Horak I.R.2, Drobot L.B.2, Matyshevska O.P.1, Prylutskyy Yu.I.1, Slobodyanik M.S.1, Ritter U.3
1Taras Shevchenko National University of Kyiv, 64 Volodymyrska Str., Kyiv, 01601, Ukraine
E-mail: psvit@bigmir.net
2Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Leontovich Str., 9, Kyiv, 01030, Ukraine
3Technical University of Ilmenau, 25 Weimarer Str., Ilmenau, 98693, Germany
Cisplatin (CP) is a drug that is used against tumors of various origins (lymphomas, cancer of breast, brain, lung etc.). In this work the cells of Lewis lung carcinoma (LLC) were used as experimental model. Since traditional antitumor drugs are characterized by cardio-, hepato-, nephro- and pulmonary toxicity, it is important to find compounds that reduce these side effects. In this sense, the carbon nanostructures, in particular C60 fullerene, are of considerable interest. C60 able to penetrate through biological membranes and to exhibit antioxidant properties. The unique structure of C60 molecule makes it possible to modify its surface with chemotherapeutic agents. The aim of this study was to create a С60-CP nanocomplex and to investigate its anticancer activity against LLC cells.
A highly stable aqueous colloid solution of C60 (10-4 M, purity >99.5%, 50 nm average size) was used in the study. A C60-CP nanocomplex was prepared by mixing the C60 colloid (0.15 mg/ml) and CP saline (0.15 mg/ml) solutions in a molar ratio of 1:2.4. The mixture was treated by ultrasound (22 kHz, 20 min) with further stirring on the magnetic stirrer (400 rpm, 18 h). This nanocomplex was characterized by physical methods such as DLS, AFM and STM.
By using the MTT test the time- and concentration-dependent decrease of LLC cells viability induced by C60-CP nanocomplex was found. Addition of nanocomplex at a final 1.5 µg/ml concentration of CP caused significant inhibition of LLC cells viability after 72 h incubation, the IC50 value for nanocomplex was found to be 6,7 times lower as compared with free CP. It was shown that nanocomplex induced the decrease of LLC cells number and changes of cells shape and size, the most of the cells were rounded and did not form sprouts, while free cisplatin at this concentration did not affect LLC cells morphology. By using the flow cytometry with propidium iodide it was shown that CP and C60-CP nanocomplex affected the cell cycle distribution of LLC cells. A significant LLC cells content was detected in G2/M phase under the action of Cis-Pt, while nanocomplex enhanced the content of cells in proapoptotic subG1 phase. The data obtained show that complexation of CP with C60 fullerene allowed to reduce IC50 index and to enhance the cytotoxic effect of this chemotherapeutic drug against LLC cells.
This work was partially supported by STCU project N 6256.