INCREASED EXPRESSION LEVEL OF ADAPTOR PROTEIN Ruk/CIN85 IN DOXORUBICIN-RESISTANT HUMAN NON-SMALL LUNG ADENOCARCINOMA MOR CELLS IS ASSOCIATED WITH THEIR METABOLIC REPROGRAMMING

Abstract

The aim of the present study was to find out the role of Ruk/CIN85 in modulation of activities/content of key enzymes/components of glycolysis and hydrogen peroxide using as a model human NSCLC MOR wild type and resistant to drugs MOR/0.2R cells. Materials and methods. MOR (ECACC 84112312) and MOR/0.2R (ECACC 96042335), drug-resistant cell line, were cultured in the complete RPMI medium under standard conditions. Enzymes activity, content of metabolites and protein in cell extracts and the conditioned cell culture medium were estimated by spectrophotometric and fluorometric assays. Results. Using RT2-PCR it was revealed that the level of Ruk/CIN85 mRNA in drug-resistant MOR cells was 10 times higher than in parental MOR cells. The activities of lysyl oxidase (LOX) and diamine oxidase (DAO) were significantly higher in resistant cells It has been shown that these enzymes are associated with aggressiveness of tumor cells. Based on the obtained results, we draw a conclusion that observed changes in the intensity of glycolysis, amine oxidases activities and content of hydrogen peroxide in doxorubicin-resistant MOR/0.2R cells positively correlate with the expression level of the adaptor protein studied. Conclusions. In conclusion, it can be assumed that the adaptor protein Ruk/CIN85 is involved in metabolome reprogramming and may function as an important component of regulatory networks required for the acquisition of drug resistant phenotype by NSCLC cells.