Lypova N. Unique alternative splice-variants of 6-phosphofructo-2-kinase/fructose-2,6-bispohsphatase-1 and -2 from animal and human cells

The object of study: mRNA of PFKFB2, PFKFB and its alternative splice-variants. Purpose: investigation of the alternative splice-variants of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-2 (PFKFB2) mRNA in mouse brain and in the rat tissues with experimental diabetes mellitus as well as PFKFB1 and PFKFB2 mRNA in glioma cells and its subline with suppressed function of ERN1 in hypoxia and glucose or glutamine deprivation. Methods: RNA and plasmid DNA isolation, cDNA synthesis, spectrophotometry, electrophoretic analysis of nucleic acids, cloning, PCR and qPCR methods, computer analysis of data. Theoretical and practical results, novelty: New alternative splice variants of PFKFB-2 mRNA were identified in rat and mice tissues. These splice variants of PFKFB-2 mRNA have different inserts and/or deletions in 6-phosphofructo-2-kinase as well as in fructose-2,6-bisphosphatase part. The results of this investigation clearly demonstrate that diabetes mellitus significantly affects the expression of PFKFB-2 mRNA and its alternative splicing in the kidney and lungs and show the complexity of regulatory mechanisms of glucose metabolism in this disease. We have identified unique alternative splice variants of PFKFB1 and PFKFB2 mRNA with deletions or insert in its 5’-region. It was shown that blockade of ERN1 enzyme function, the key endoplasmic reticulum stress sensor, increases the expression levels of PFKFB1 and PFKFB2 mRNAs as well as its alternative splice variants. The dynamics of mRNA PFKFB2 expression during the day was investigated. Field of application and performance: alternative splicing pre-mRNA of PFKFB2 and 1 can be used as target for the development of new strategies of anticancer drugs and treatment of metabolitic and proliferative disorders.