Danilovskyi S. Regulation of the expression of TP53 and its dependent proteins in glioma cells line U87

Object: the function of genes TP53, MDM2, PERP, USP7, RBL1, CSNK2, NME6, TOPORS, ZMAT3, TP53BP1 and TP53BP2. Objective: - study of the dependence of expression of the genes TP53, MDM2, PERP, USP7, RBL1, CSNK2, NME6, TOPORS, ZMAT3, TP53BP1 and TP53BP2 the functional activity of a signaling enzyme of endoplasmic reticulum stress ERN1 in conditions of hypoxia and glutamine or glucose deficiency in the cells of glioma lines U87. Methods: glioma cell line U87 and its sublines cultivation, total RNA extraction, agarose electrophoresis of nucleic acids, reverse transcription, polymerase chain reaction, quantitative polymerase chain reaction, extraction of cytosol and nucleus protein fractions from the cell cultures, precipitation of proteins from the cell culture media, spectrophotometric identification of protein amounts, protein polyacrylamide electrophoresis, immunoblotting, computer analysis of polymerase chain reaction results, statistical methods of data processing. It was shown for the first time that the expression of TP53, MDM2, PERP, USP7, RBL1, CSNK2, NME6, TOPORS, ZMAT3, TP53BP1 and TP53BP2 genes in the glioma cell depended on functional activity of ERN1 and revealed that changes in the expression of the studied genes correlated with the decreasing of proliferation of cells with suppressed ERN1 activity. It was established that changes in the expression of TP53, MDM2 and USP7 genes in glioma cells under condition of ERN1 inhibition were caused by suppression of kinase, but not endoribonuclease activity of the ERN1 and changes in the expression of PERP gene - by both activities.