Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma

Abstract

Background: The methylation of Ubiquitin Carboxyl Terminal Hydrolase 1 (UCHL1) gene’s promoter has been reported as the etiological factor of nasopharyngeal tumorigenesis. Purpose: This study is designed to establish a protocol for detecting methylation of UCHL1 gene’s promoter in nasopharyngeal carcinoma (NPC) in a Vietnamese population. Materials and methods: 10 samples of NPC biopsy tissues and 10 samples of non-cancerous swabs were collected from the local hospital. Chloroform/Phenol method and Nested-MSP assays were established to detect methylation of a target gene. Results: The isolated DNA reached purity and high concentration which were confirmed by the method of absorbance measurement at 260nm and 280nm. Additionally, the Nested-MSP products of methylation or unmethylation were analyzed and visualized in the agarose gel with the band of 169bps and 210bps, respectively. By sequencing, it was confirmed that the two sets of primer could distinguish the status of methylation and unmethylation of UCHL1 gene’s promoter. Conclusion: Our data suggested that the current protocol could successfully identify the status of methylation and/or unmethylation of UCHL1 gene’s promoter.

Keywords

• epigenetics
• methylation
• nested-msp
• ubiquitin carboxyl terminal hydrolase 1
• unmethylation