Molecular analysis of the effects of histon methyl transferases in the treatment of SH-SY5Y human neuroblastoma cells formed with alzheimer's disease model

Alzheimer's disease (AD) is a neurodegenerative disease that is seen in people over 65 years old and is characterized by deterioration in behavioral and cognitive functions such as memory, comprehension, language and reasoning. AD is characterized by the accumulation of abnormal neuritic plaques called amyloid beta 42 (Aβ42) and neurofibrillary tangles in the brain. Although AD can be inherited as an autosomal dominant disease associated with mutations in some genes, most of these genes are under the influence of epigenetic mechanisms such as DNA and histone modifications. Because epigenetic modifications are reversible, they can potentially be targeted by pharmacological intervention. Accordingly, epigenetic drugs may be useful in the treatment of important diseases such as cancer, cardiovascular disorders, and brain disorders. In this context, it was aimed to develop therapeutic approaches that can positively affect the prognosis of the disease by using methyl transferase enzyme inhibitors that will change the expression levels of genes involved in the increase of Aβ plaques by creating an Alzheimer's model with sufficient Aβ42 expression in SH-SY5Y human neuroblastoma cells in this study. For this purpose, after culturing SH-SY5Y cells under appropriate conditions, the Alzheimer's model was created with the Aβ42 protein application, and quantitative measurements of Aβ protein, which started to be produced in cells, were performed by the ELISA method. In addition, the expression levels of genes associated with Aβ production were measured by Real Time-Polymerase Chain Reaction (RT-PCR) and the Alzheimer model formed in the cells was controlled at the gene level. By applying methyltransferase inhibitors to the cells at different concentrations such as Azacitidine, Deazaneplanocin and SGC0946, which will reduce the amount of amyloid beta plaques formed, optimum concentrations were determined with the MTT test and the amount of Aβ42 in Alzheimer's model cells is measured at the protein level by ELISA method, as well as the expression levels of genes such as APP, PSEN, BACE1, GSAP, BDNF associated with the disease determined by RT-PCR quantitatively. As a result of the study, a statistically significant decrease was found in Aβ42 plaque levels with the effect of methyltransferase inhibitors used. Significant changes were detected in the expression levels of genes associated with Aβ42 plaques at different rates after each chemical used, which could positively affect the prognosis of the disease. As a result, it has been observed that the chemicals used cause significant changes in cell viability by changing the existing epigenetic modifications in the cells.