Acta Medica Iranica 2018. 56(1):21-27.

Promoter Hypermethylation of the Eyes Absent 4 Gene is a Tumor-Specific Epigenetic Biomarker in Iranian Colorectal Cancer Patients
Matineh Barati Bagerabad, Sahar Tavakolian, Mohammad Reza Abbaszadegan, Mohammad Amin Kerachian


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.


DNA methylation; Colorectal cancer; EYA4; Screening

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Coppedè F. Epigenetic biomarkers of colorectal cancer: Focus on DNA methylation. Cancer Letters. 2014;342:238-247.

Hessami Arani S, Kerachian MA. Rising rates of colorectal cancer among younger Iranians: is diet to blame? Curr Oncol 2017;24:e131-7.

. Shen L, Toyota M, Kondo Y, et al. Minoru Toyota et al. Integrated genetic and epigenetic analysis identifies three different subclasses of colon cancer. PNAS. 2007;104:18654-9.

. Coppedè F, A Lopomo, R Spisni, & L Migliore. Genetic and epigenetic biomarkers for diagnosis, prognosis and treatment of colorectal cancer. World J Gastroenterol 2014;20:943-956.

Ashktorab H & Brim H. DNA Methylation and Colorectal Cancer. Curr Colorectal Cancer Rep. 2014;10:425-430.

Imperiale TF, Ransohoff DF, Itzkowitz SH, et al. Multitarget Stool DNA Testing for Colorectal-Cancer Screening. N Engl J Med. 2014;370:1287-1297.

Lao VV and Grady WM. Epigenetics and Colorectal Cancer. Nat Rev Gastroenterol Hepatol. 2011;8:686-700.

Lange CP & Laird PW. Clinical applications of DNA methylation biomarkers in colorectal cancer. Epigenomics 2013;5:105-108.

Shirahata A, Sakata M, Sakuraba K, et al. Vimentin Methylation as a Marker for Advanced Colorectal Carcinoma. ANTICANCER RESEARCH. 2009;29:279-282.

Willyard C. To foster screening, new colon cancer tests emphasize convenience. NATURE MEDICINE. 2014;20:322-3.

Borsani G, DeGrandi A, Ballabio A, et al. EYA4, a novel vertebrate gene related to Drosophila eyes absent. Hum Mol Genet. 1999;8:11-23.

Tadjuidje E and Hegde RS. The Eyes Absent Proteins in Development and Disease. Cell Mol Life Sci. 2013;70:1897-1913.

Zou H, Osborn NK, Harrington J, et al. Frequent Methylation of Eyes Absent 4 Gene in Barrett's Esophagus and Esophageal Adenocarcinoma. Cancer Epidemiol Biomarkers Prev. 2015;14: 830-834.

Kim SJ, Tae CH, Hong SN, et al. EYA4 Acts as a New Tumor Suppressor Gene in Colorectal Cancer. Mol Carcinog 2015;54:1748-57.

Wilson IM, Vucic EA, Enfield KSS, et al. EYA4 is inactivated biallelically at a high frequency in sporadic lung cancer and is associated with familial lung cancer risk. Oncogene. 2014;33: 4464-4473.

Oster B, Thorsen K, Lamy P, et al. Identification and validation of highly frequent CpG island hypermethylation in colorectal adenomas and carcinomas. Int. J. Cancer. 2011;129:2855-2866.

Osborn NK, Zou H, Molina J, et al. Aberrant Methylation of the Eyes Absent 4 Gene in Ulcerative Colitis–Associated Dysplasia. CLINICAL GASTROENTEROLOGY AND HEPATOLOGY. 2006;4:212-218

Mitchell SM, Ross JP, Drew HR, et al. A panel of genes methylated with high frequency in colorectal cancer. BMC Cancer. 2014;14:54.

Xu P-X, Adams J, Peters H, et al. Eya1-deficient mice lack ears and kidneys and show abnormal apoptosis of organ primordia. Nat Genet. 1999;23:113-117.

Xu P-X, Woo I, Her H, Beier DR, Maas RL. Mouse Eya homologues of the Drosophila eyes absent gene require Pax6 for expression in lens and nasal placode. Development. 1997;124:219 -231.

Abdelhak S, Kalatzis V, Heilig R, et al. Human homologue of the Drosophila eyes absent gene underlies branchio-oto-renal (BOR) syndrome and identifies a novel gene family. Nat Genet. 1997;15:157-164.

Abdelhak SL, Kalatzis V, Heilig R, et al. Clustering of mutations responsible for branchio-oto-renal (BOR) syndrome in the eyes absent homologous region (eyaHR) of EYA1. Hum Mol Genet. 1997;6:2247-55.

Vincent CL, Kalatzis V, Abdelhak S, et al. BOR and BO syndromes are allelic defects of EYA1. Eur J Hum Genet. 1997;5:242-6.

Wayne S, Robertson NG, DeClau F, et al. Mutations in the transcriptional activator EYA4 cause late-onset deafness at the DFNA10 locus. Hum Mol Genet. 2001;10:195-200.

Pfister M, Toth T, Thiele H, et al. A 4-bp insertion in the eya-homologous region (eyaHR) of EYA4 causes hearing impairment in a Hungarian family linked to DFNA10. Mol Med. 2002;8:607-11.

Kisiel JB, Yab TC, Nazer Hussain FT, et al. Stool DNA testing for the detection of colorectal neoplasia in patients with inflammatory bowel disease. Aliment Pharmacol Ther. 2013;37:546-54.

Jones PA, & Baylin SB. The fundamental role of epigenetic events in cancer. Nat Rev Genet. 2002;3:415-28.

Jones PA & Laird PW. Cancer epigenetics comes of age. Nat Genet. 1999;21:163-7.


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