Methylation Analysis of OCT 4 Gene in AML and ALL Patients Using Sodium Bisulfite

Abstract

Methylation is among the most important epigenetic modifications. This modification is involved in gene silencing and activation of certain genes. OCT 4 is a progenitor gene that encodes the OCT 4 transcription factor, which is essential for the pluripotency and self-renewal of embryonic stem cells. This gene is only active during pre-embryonic stages of development, when cells are differentiating, and becomes inactive by certain epigenetic modifications, such as methylation, when cells are differentiated. Removal of methylation from the regulatory region of this gene causes disruption of normal cellular differentiation and ultimately leads to an imbalance. This imbalance leads to leukemia. This study was designed to analyze the relationship between the methylation pattern of OCT4 in AML and ALL patients. Forty samples each of control, AML, and ALL were collected from PIMS Islamabad. Identification was performed using TLC count and bone marrow biopsy. Gel electrophoresis was performed on the extracted DNA, and Genomic DNA was quantified. A conventional method was used for bisulfite genomic treatment. Primers were designed, and PCR was performed. The methylation profile of OCT 4 was analyzed in 40 samples each of AML, ALL, and control samples. From the collected clinical data from patients, it was observed that the average WBCs count in AML and ALL patients was significantly higher than normal. From the data, it can be concluded that males are more prone to AML and ALL than females. It was found that OCT4 was hypomethylated in most AML and ALL patients. In AML and ALL patients, hypomethylation status of the OCT4 gene was observed after bisulfite genomic treatment when amplified with conventional and bisulfite-specific primers, and the results of amplification confirmed the conversion of unmethylated cytosine to uracil.