Mohd. Ridah, Lailatul Jalilah and Ismail, Aziah and A. Talib, Norlelawati and Muhammad, Siti Aeshah @ Naznin and Hussain, Faezahtul Arbaiyah and Zainuddin, Norafiza
(2016)
Pyrosequencing-based quantitative identification of p16 methylation in diffuse large B-cell lymphoma at two centres in the East Coast of Malaysia.
Journal of Biomedical and Clinical Sciences, 1 (1).
pp. 12-17.
E-ISSN 2550-147X
Abstract
Introduction: Methylation of promoter region of p16 leading to gene silencing has been implicated in a wide range of malignancies including lymphomas. In diffuse large B cell lymphoma (DLBCL) particularly, a varying
percentage of epigenetic inactivation of p16 promoter region was observed ranging from 16 - 54%. However, quantitative analysis of p16 promoter methylation in DLBCL has not been extensively studied in Malaysia. Objective: This study aims to quantitatively analyse p16 methylation in DLBCL samples using pyrosequencing technique. Methods: Genomic DNA was extracted from 16 formalin-fixed paraffin-embedded lymphoma tissue blocks from patients diagnosed with DLBCL. Samples were retrieved from Hospital Tengku Ampuan Afzan, Pahang and Hospital Universiti Sains Malaysia, Kelantan. Primers were designed to amplify bisulfite-treated DNA targeting p16 promoter region. Methylation status of 7 CpG sites was determined by pyrosequencing. Results: All the 16 samples studied showed promoter methylation of p16. The
range of mean methylation percentage was between 18 to 81%. Conclusion: The present study has successfully measured the level of methylation of p16 in all 7 CpG sites despite the limitation in sample size. Since p16 methylation is a common event in our series of DLBCL cases, it is worth including a larger sample size in future studies to increase the chance of finding a significant correlation with clinical parameters.
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