The effect of erythropoietin on MRMA level in shsy-5y cell line by microarray

The Effect of Erythropoietin on mRNA Level in SHSY-5Y Cell Line by Microarray Zeynep Filiz ÖNKAL, Dokuz Eylul University The Institute of Health Sciences, Department of Neuroscience, 35340 Inciraltı, IzmirErythropoietin (EPO) is a hematopoietic growth factor and cytokine which stimulates erythropoiesis. In recent years it?s been shown that EPO and its receptor is expressed in the brain, neurons and the glial cells. EPO has a critical role in the development, maintenance, protection and repairment of the nervous system. A possible mechanism of EPO is gene regulation at the transcriptional level. In this study, our aim is to investigate the effects of EPO on mRNA expression levels in SHSY-5Y neuroblastoma cell line. In our project, the transcriptional effects of EPO was examinated by microarray. For this purpose neuronal cell line SHSY-5Y has been induced by EPO and any mRNA expression changes that occured has been detected by microarray and Quantitative Polymerase Chain Reaction (qPCR) methods both.According to our results, by eBayes hypothesis test, there is no significant changes in control versus 24 hour EPO induction group. Our results show that in comparision control group versus 48 hours EPO induction group, 438 gene upregulated and 482 genes down regulated. <0,5 fold change determined as decreased gene expression level and > 1,5 was determined as increased gene expression level. According to this, 52 gene fold change >1,5 and 23 genes fold changes <0,5. We choose 26 genes for qPCR confirmation depend on literature. The concordance between microarray and qPCR results was found. Our results show that 18 gene expression was significantly increase and 8 genes expression was significantly decrease after 48 hour EPO induction. Using KEGG and reactome applications, we investigated releated pathways such as extracellular matrix organisation, axon guidance, signal transduction, protein metabolism. Key Words: Erythropoietin, SHSY-5Y Cell line, Gene expression, Microarray