Abstract 4037: Downregulation of ZNF516 and INTS1 is related with promoter methylation in cervical uterine cancer cell lines

Abstract

Abstract Introduction: One of the most common neoplasia in developing countries is cervical uterine carcinoma (CC). HPV infection is recognized as the major risk factor for carcinogenesis of cervix. However, not all infected women will develop a cervical cancer. Several studies have proposed DNA methylation as early biomarker in CC. A higher promoter methylation is associated with lower gene expression. Aim: To evaluate the association between DNA methylation and gene expression of ZNF516 and INTS1 in cervical cancer cell lines. Materials and Methods: DNA isolated from 12 normal and 7 cervical cancer tissue samples was enriched by Methylated DNA Immunoprecipitation (MeDIP) and hybridized to Nimblegen 385K CpG Islands plus Promoter arrays (MeDIP-chip). Bioinformatics strategies were used for background correction, array normalization and data analysis of differentially methylated genomic regions by Methylation Specific PCR in a cohort of 221 normal, premalignant, and cervical cancer samples. Relative expression analysis was performed in a normal cervical epithelium cell line (ECT1 E6/E7) and three cervical cancer cell lines (C-4I, SiHa and C-33A) using real time PCR. MSP was also used to examine promoter methylation in normal cervical and cervical cancer cells lines. Results: ZNF516 and INTS1 were differential methylated in the promoter region of normal and cervical cancer samples in the MeDIP-chip discovery project. The methylation frequency was 100% in cervical cancer cell lines. Relative expression of ZNF516 was significantly lower in C-4I and SiHa (p <0.05) in relation with the normal cell line. C-33A showed elevated promoter methylation and a low expression of ZNF516, however, this result was not significant. INTS1 expression was significantly lower in C-33A and SiHa (p <0.05) in relation with the normal cell line. C-4I showed a high promoter methylation and a low expression of INTS1, however, this result was not significant. Conclusions: We have identified ZNF516 and INTS1 as potentials biomarkers for cervical cancer. These genes may be a useful tool for diagnosis and clinical management of cervical cancer. These results need to be confirmed in clinical samples, from an independent laboratory before the gene panel is examined in a Phase II Biomarker Development Trial. Work sponsored by CORFO Project 09CN14-5960 and Postdoctoral Fondecyt Research Project 3120141. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4037. doi:1538-7445.AM2012-4037