Qualitative detection of p16INK4a tumor suppressor gene methylation in diffuse large b-cell lymphoma using methylation-specific PCR

• Main Authors: Mohd. Ridah, Lailatul Jalilah, Md Dzali, Nurul Balqis, Zainuddin, Norafiza
• Format: Conference or Workshop Item
• Language: English; English
• Published: 2014
• Subjects: RC0254 Neoplasms. Tumors. Oncology (including Cancer)
• Online Access: http://irep.iium.edu.my/43663/1/KAHS_RESEARCH_WEEK_2014_ABSTRACT_BOOK.pdf; http://irep.iium.edu.my/43663/4/43663.pdf; http://irep.iium.edu.my/43663/; http://www.iium.edu.my/kahs/events/kahs-research-week-2014

INTRODUCTION: Diffuse large B-cell lymphoma (DLBCL) is an aggressive malignancy of non-Hodgkin lymphoma, mostly involve lymph nodes. It is the most common lymphoma among adult with median age 70 years. p16 is a tumour suppressor gene (TSG) which inhibits cyclin-dependent kinase thus inactivates Rb protein and blockade G1 phase. Silence of p16 induced by DNA methylation epigenetically has been reported as one of the factors toward DLBCL occurrence with varying percentages. Loss of p16 protein causes unregulated cell division which may lead to cancer. In this study, we aim to identify the presence of P16INK4a gene methylation in DLBCL and to correlate the methylation status with patients’ age and gender. MATERIALS AND METHODS: The samples of DLBCL were retrieved from two study groups; Kelantan and Pahang states. Methylation-specific PCR (MSP) was utilized to detect p16 methylation using specific primers in 88 formalin-fixed paraffin embedded DLBCL tissues including five normal thyroid samples. The PCR products were then visualized on 2% agarose gels under UV illumination. RESULTS AND DISCUSSION: 64 of 88 (73%) samples were found to have p16 methylation. There was no p16 methylation spotted in five normal thyroid samples. We found statistically significant association between p16 methylation and patients’ age (>50 years) with p = 0.023. While, there is no association between p16 methylation and gender observed in this study. CONCLUSION: p16 gene methylation has been detected in most of the samples with 72% of methylation occurred among patients more than 50 years. This could provide data of gene methylation in DLBCL among Malaysian. Thus, more studies should be carried out for further investigation.