Methylation Status Of P15 Gene In Esophageal Squamous Cell Carcinoma Using Methylation-Specific PCR

Document Type : Original Article

Authors

CLINICAL BIOCHEMISTRY DEPARTMENT, SCHOOL OF MEDICAL SCIENCES, TARBIAT MODARRES UNIVERSITY, TEHRAN, IRAN

Abstract

Introduction: Promoter methylation has an inhibitory effect on gene expression. Thus we investigated methylation patterns of p15 gene in Esophageal Squamous Cell Carcinoma (ESCC), one of the most frequent and fetal cancers in the world.
Material and methods: p15 promoter methylation status of 30 ESCC and 10 normal specimens was investigated using methylation specific PCR. DNA extracted from the specimens was digested by restriction enzyme. The digested DNA was modified by sodium bisulfite. Using methylated and unmethylated specific primers, methylation status was determined based on the presence of the methylated and unmethylated specific PCR products. Also DNA extracted from normal blood and CpG methylase (SssI)–treated genomic DNA were used as the negative and positive controls respectively.
Results: Our results showed that all normal specimens were unmethyalted and only 5 out of 30 tumor samples (16.6%) were methylated. Statistical analysis (Fisher’s exact tests) showed no significant correlation between p15 gene promoter methylation and ESCC cancer.
Conclusion: According to the results of the study, p15 gene does not play a significant role in aberrant methylation process in ESCC, other mechanisms may involve in p15 inactivation during ESCC development. However, due to limited number of specimens, we recommend further investigation using a larger sample.

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