Study of the promoter methylation status of RASSF1A gene in oral squamous cell carcinoma

Document Type : Original Article

Authors

1 Assistant Lecturer of Oral and Dental Pathology Department, Faculty of Dental Medicine, Assiut, AL-Azhar University, Egypt.

2 Professor of Oral and Dental Pathology Department, Faculty of Dental Medicine, (Boys-Cairo), Al-Azhar University, Egypt.

3 Assistant Professor of Clinical Pathology Department, South Egypt Cancer Institute, Assiut University, Egypt.

4 Professor of Oral and Dental Pathology Department, Faculty of Dental Medicine, Assiut, Al-Azhar University, Egypt.

Abstract

Oral cancer is one of the major global health problems. Methylation is one of the epigenetic modifications that play an important role in the transcriptional inactivation of tumor suppressor genes in human cancer. Aim: The present study was directed to assess the Ras-association domain family 1A (RASSF1 A) gene promoter methylation status in oral squamous cell carcinoma (OSCC) and its correlation to patient’s clinicopathological data. Material and methods: Formalin-fixed, paraffin-embedded tissue blocks of fifty patients with histopathologically confirmed OSCC in addition to 10 normal oral mucosa blocks were included in this study. Hematoxylin and Eosin (H&E) stain was used to confirm the diagnosis of cases as 16 well differentiated, 19 moderately differentiated and 15 poorly differentiated OSCC. Sections of each block were deparaffinized and prepared by DNA extraction for detection of RASSF1A gene methylation status by methylation-specific polymerase chain reaction (MSP) technique. Results: RASSF1A promoter methylation was not detected in normal cases, but it was found in 30% (15/50) of OSCC cases. The increase of methylation in OSCC was significant (p-value=0.045). The increase of methylation in poorly differentiated OSCC (6/15, 40%) was more than that.0 in moderately differentiated OSCC (7/19 36.8%) but both were significante in relation to normal mucosa (p-values= 0.021, 0.027 respectively). Well-differentiated OSCC was the lowest in methylation (2/16, 12.5%), while its increase than normal was not significant (p-value= 0.244). There was direct correlation between methylation and the lymph nodes metastasis. Conclusion: RASSF1A promoter methylation significantly associated with OSCC, the decrease of cell differentiation in histopathological stages and lymph node metastasis. Hence, RASSF1A promoter methylation could be used as a tumor marker in addition to histopathological stage and lymph nodes metastasis predictor in OSCC.

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