Karbovskyi L. The expression of circadian and retinoblastoma related genes in ERN1-deficient glioma cells

3. The object of study: human CSNK1, CSNK2, PER1, PER2, PER3, CLOCK, BMAL1, CRY1, DEC2, RB1, RBL1, RBAP48, RBAP46, KDM5A, JARID1B, E2F1, E2F3, RNF40, EID1 and CTIP genes. Purpose: study of the expression of circadian, casein kinases 1 and 2, retinoblastoma and retinoblastoma-related genes in ERN-deficient glioma U87 cells in hypoxic, glutamine or glucose deprivation conditions. Methods: RNA extraction, spectrophotometry, nucleic acid electrophoresis, cDNA synthesis, chromatin immunoprecipitation, polymerase chain reaction methods. Theoretical and practical results, novelty: It was shown that the blockade of ERN1 leads to an increase in the expression levels of PER1, PER3 and CLOCK mRNA; while CRY1, PER2, BMAL1, BMAL2 and DEC2 mRNA levels are decreased. Moreover, the expression levels for most of studied genes are increased under glucose or glutamine deprivation conditions both in control and ERN1-deficient glioma cells. However, ERN1 knockdown modified the effect of these ischemic conditions. We have demonstrated that the expression of most retinoblastoma-related genes is dependent on the endoplasmic reticulum - nuclei-1 signaling enzyme function in normal, hypoxic and glucose or glutamine deprivation conditions. Field of application and performance: studied mechanisms of gene expression regulation will contribute to find perspective target genes for development new strategies of tumor growth suppression.