Promoter hypermethylation and downregulation of the FAS gene may be involved in colorectal carcinogenesis

Manoochehri,Mehdi; Borhani,Nasim; Karbasi,Ashraf; Koochaki,Ameneh; Kazemi,Bahram

doi:10.3892/ol.2016.4578

Promoter hypermethylation and downregulation of the FAS gene may be involved in colorectal carcinogenesis

Authors:
- Mehdi Manoochehri
- Nasim Borhani
- Ashraf Karbasi
- Ameneh Koochaki
- Bahram Kazemi
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Affiliations: Biotechnology Department, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran 19857‑17453, Iran, School of Medicine, Shahroud University of Medical Sciences, Shahroud 36247‑73955, Iran, Gastroenterology and Hepatology Department, Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran 14359‑15371, Iran, Cell and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran 19857‑17453, Iran
Published online on: May 16, 2016 https://doi.org/10.3892/ol.2016.4578
Pages: 285-290

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Abstract

Aberrant DNA methylation has been investigated in carcinogenesis and as biomarker for the early detection of colorectal cancer (CRC). The present study aimed to define the methylation status in the regulatory elements of two proapoptotic genes, Fas cell surface death receptor (FAS) and BCL2‑associated X protein (BAX). DNA methylation analysis was performed in tumor and adjacent normal tissue using HpaII/MspI restriction digestion and methylation‑specific polymerase chain reaction (PCR). The results observed downregulation of the FAS and BAX genes in the CRC tissues compared with the adjacent normal samples. Furthermore, demethylation using 5‑aza‑2'‑deoxycytidine treatment followed by reverse-transcription quantitative PCR were performed on the HT‑29 cell line to measure BAX and FAS mRNA expression following demethylation. The 5‑aza‑2'‑deoxycytidine treatment resulted in significant FAS gene upregulation in the HT‑29 cell line, but no significant difference in BAX expression. Furthermore, analysis of CpG islands in the FAS gene promoter revealed that the FAS promoter was significantly hypermethylated in 53.3% of tumor tissues compared with adjacent normal samples. Taken together, the results indicate that decreased expression of the FAS gene due to hypermethylation of its promoter may lead to apoptotic resistance, and acts as an important step during colorectal carcinogenesis.

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