Background: Epstein-Barr virus (EBV) has been classified among group 1 listof infectious agentsthat is carcinogenic to humans. Some primary colonic cancers are often overexpressing the p16 protein while in contrast; others showed the high incidence of p16 repression by methylation. Objective: The present study was conducted to explore and compare the demo-pathological relation of EBV infection along with p16 expression in normal colonic mucosa, benign colon tumors, and colonic denocarcinoma. Patients and Methods: Seventy (70) colonic mucosal biopsies were enrolled and examined for EBERs and P16 gene expression which were done by chromogenic in situ hybridization (CISH) and immunohistochemical technique (IHC), respectively. These samples were belonged to (30) patients diagnosed with colorectal cancer mass, (25) from benign colon tumors (15) control tissues, which were proved by colonoscopy and histopathological examinations as apparently normal colorectal tissues. Results: Malignant colonic tumors showed positive results of EBERs -CISH detection in 56.7% (17 out of 30 tissues) while in benign colonic tumors were 24 % (6 out of 25 tissues), followed by the apparently healthy colonic control tissues were 20% (3 out of 15 tissues). The present positive p16-IHC results in malignant colonic tumors were 60% (18 out of 30 tissues) while in the benign colonic tumor and apparently healthy colon control tissues were 68% (17 out of 25 tissues) and 20% (3 out of 15 tissues), respectively. Conclusion: it was concluded from this study that the high rates ofEBV- EBERs expression could point to a their possible roles incolonic carcinogenesis, meanwhile, the p16- proteinexpression in colonic adenocarcinoma as compared to benign colon tumors and normal colonic mucosae could played roles in struggling against both EBV infection as well as colonic carcinogenesis.