Nanochemistry and biotechnology

Cytotoxic and apoptotic effects of сis-Palladium(II)3-(2-pyridyl)-5-methyl-1,2,4-triazole in combination with C60 fullerene 
against LLC cells

Grynyuk I.I.1, Prylutska S.V.1, Skaterna T.D.2, Horak I.R.2, Raspertova I.V.1, Doroshchuk R.O.1, Matyshevska O.P.1, Drobot L.B.2, Slobodyanik M.S.1, Ritter U.3
1Taras Shevchenko National University of Kyiv, 64 Volodymyrska Str., Kyiv, 01601, Ukraine
E-mail: igrynyuk@ukr.net
2Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 9 Leontovicha Str, Kyiv, 01030, Ukraine
3Technical University of Ilmenau, 25 Weimarer Str., Ilmenau, 98693, Germany
Lung cancer is a prevailing disease with a high mortality. Cisplatin and its analogs are effective agents for cancer treatment, however, their application is limited by drug resistance development and severe nephro-, oto-, neuro- and hematological toxicity. Recent studies  are mainly focused on the search for new coordination platinum metal complexes and their combinations with nanostructures for reducing drug toxicity and improving its therapeutic properties. Since Palladium has many chemical similarities with platinum a new сis-Palladium(II) 3-(2-pyridyl)-5-methyl-1,2,4-triazole complex (сis-Pd complex) was synthesized. A representative of carbon nanostructures C60 fullerene was used as a possible modulator of сis-Pd complex biological action.
The aim of this study was to estimate the effect of cis-Pd complex on the viability, morphology and cell cycle distribution of LLC (Lewis lung cancer) cells separately and in combination with C60 fullerene. 
Cis-Pd complex was synthesized and characterized using NMR and FTIR spectroscopy (the purity of compound was ≥98%). A highly stable water colloid solution of C60 fullerene (10-4 M, purity >99.5%, nanoparticles of 50 nm average size) was synthesized at Technical University of Ilmenau (Germany). 
LLC cells viability was estimated using MТТ test at 24, 48 аnd 72 h after addition of cis-Pd complex in 0.2 - 10 µМ concentration range. The cytotoxic dose- and time dependent effect of cis-Pd complex was shown. The cytotoxic activity of cis-Pd complex against LLC cells at 48h incubation was found to be higher as compared with cisplatin, IC50 value for cis-Pd complex was 2.744 μM, whereas for cisplatin - 27.79 μM. Both morphological changes and significant decrease of LLC cells number were detected at combined action of cis-Pd complex and C60 fullerene. Cell cycle analysis revealed the increased LCC cells content in G2/M phase after treatment with cis-Pd complex and the  increased cells content in apoptotic subG1 phase after combined treatment with cis-Pd complex and C60 fullerene.
Thus, cis-Pd complex showed a pronounced cytotoxic effect, which was intensified at combined treatment with C60 fullerene.