Functional analysis of genes may involved in alzheimer disease

Ferroptosis is a newly discovered form of iron-dependent death triggered by lipid peroxides. Many pathways and related genes have been identified for regulating ferroptosis. Some of these genes have been associated with various disease pathogenesis. Alzheimer's disease (AD) is a neurodegenerative disease characterized by dementia and its prevalence has increased in recent years. The mechanism of cell death in AD pathogenesis is not fully understood. In this study, artificial ferroptosis was created with the ferroptosis inducer erastin and its inhibitor ferrostatin-1 in HEPG2 cell line. Cell counts using hemocytometer and flow cytometry experiments were performed to confirm ferroptosis. After optimization of cell culture, the effect of genes (BNIP3L, TM2D1 and RAD21) thought to be associated with AD in the HEPG2 cell line was examined by quantitative RT-PCR (qPCR). At the same time, changes in expression levels of ferroptosis-related genes (GPX4 and SLC7A11) were evaluated by quantitative RT-PCR in 25 Alzheimer's patients and 21 controls. In this context, qRCR results from cell culture showed that the changes in gene expression of interest were either in the same direction only under erastin and in conditions where erastin and ferrostatin were added at the same time. It was determined that there was no significant change in qRZR results of the related ferroptosis genes in patient groups. As a result, ferroptosis was successfully induced in cell culture with erastin and ferrostatin-1. However, no significant change was observed in related gene expression. Key Words: ferroptosis, erastin, ferrostatin-1, Alzheimer's Disease, cell death