Biopolymers and cell. 2010; 26 (2): 153 - 159
RNA interference of genes of ubiquitn and proteasome 7 subunit in cardiomyocytes culture
V. O. Kyrychenko, V. S. Nagibin, L. V. Tumanovska, V. K. Rybalchenko1, V. E. Dosenko, O. O. Moibenko
Bogomoletz Institute of Physiology of NAS of Ukraine 4, Bogomoletz str., Kiev, Ukraine, 01024
1National Taras Shevchenko University of Kyiv 64, Volodymyrska Str, Kyiv, Ukraine 01033
Aim. Ubiquitn (UBB) and proteasome 7 subunit (PSMB7) genes silencing is interesting due to the crucial role of ubiquitin-dependent proteasomal system (UPS) in turnover of functional proteins, which controls program cell death (apoptosis, auophagy). Methods. We used methods of RNA interference (for UBB and PSMB7 genes silencing), fluorescence microscopy and real-time PCR. Results. It was shown that small interference RNA injection in cell culture decreased ubiquitin expression in 2,4 (P < < 0.05), PSMB7 – in 1,6 times (P > 0.05). At the same time, there was augmented level of necrotic cells with no change in number of apoptotic cells. Cells with signs of autophagy were much augmented as consequence of UPS impairment and intracellular proteins accumulation, which degrade in lysosomes. Conclusions. The data obtained testify that UBB and PSMB7 genes silencing induces cell necrosis and autophagy without changing number of apoptotic cells.
Keywords: ubiquitin, proteasome, RNA-interference, cardiomyocytes.