Tehran University of Medical Sciences Acta Medica Iranica 0044-6025 56 1 2018 02 05 21 27 EN Matineh Barati Bagerabad Medical Genetics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. AND Department of Biology, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran. Sahar Tavakolian Medical Genetics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. AND Department of Biology, University of Sistan and Balouchestan, Zahedan, Iran. Mohammad Reza Abbaszadegan Medical Genetics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. AND Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. AND Department of Human Genetics, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran. Mohammad Amin Kerachian Medical Genetics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. AND Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. AND Cancer Genetics Research Unit, Reza Radiotherapy and Oncology Center, Mashhad, Iran. 2017 03 16 2017 12 12 The aim of this study was to investigate whether hypermethylation of Eyes Absent 4 (EYA4) is also implicated in Iranian Colorectal Cancer (CRC) patients or not. From fresh frozen tissues, samples from 38 paired (cancer and normal) CRC tissue specimens were used in this study, the DNA was isolated, sodium bisulfite treated and analyzed by methylation-specific polymerase (MSP) chain reaction using primers specific for unmethylated or methylated promoter sequences of the EYA4 gene. We also analyzed EYA4 mRNA expression using real time RT-PCR. Demographic characteristics of these patients including age, sex, tumor grade, location, stage, and TNM classification were evaluated and the relationship between methylation status of the gene and clinicopathological features was analyzed. Current study indicated that EYA4 promoter hypermethylation has a sensitivity of 81.57% and specificity of 78.94%. Findings showed lower expression of EYA-4 in methylated samples in comparison with its normal adjacent tissue, although it was not significant (P>0.05). No significant associations were observed between EYA4 hypermethylation and the clinicopathological characteristics. Although the clinical patient outcome of our 38 CRC patients was not associated with EYA4 promoter hypermethylation, the high frequency of this methylation and its high sensitivity and specificity to neoplastic cells may qualify EYA4 promoter methylation as a potential candidate screening marker in Iranian population and may help to improve early detection of CRC. https://acta.tums.ac.ir/index.php/acta/article/view/6312 https://acta.tums.ac.ir/index.php/acta/article/download/6312/5039