DNA Methylation Alterations in Benzene-Exposed Gasoline Station Workers: Insights from Singleplex MethyLight Analysis

Background & Objective: Alterations in global DNA methylation levels and repetitive elements are commonly observed in various cancers and in response to environmental pollutants, particularly petroleum products such as benzene. Long-term benzene exposure is harmful to human health and is associated with an increased risk of hematological malignancies. This study aimed to investigate the association between chronic occupational exposure to benzene and DNA methylation status in TGFβ2, MAGE-A1, and LINE-1 repetitive elements among gasoline station workers. Material and Methods: This case–control study included 30 male gasoline station workers and 30 male controls. Genomic DNA was extracted from peripheral blood samples using the QIAamp DNA Mini Kit (QIAGEN) and treated with sodium bisulfite via the EpiTect Fast DNA Bisulfite Kit. Singleplex MethyLight PCR was performed to assess methylation status of MAGE-A1, TGFβ2, and LINE-1. Results: Significant differences were observed in working hours (p = 0.004) and direct benzene exposure (p < 0.0001) between workers and controls. Percent MethyLight Reference (PMR) values were significantly different between the two groups for LINE-1 (p < 0.0001), TGFβ2 (p = 0.0194), and MAGE-A1 (p < 0.0001). Conclusion: Chronic benzene exposure is associated with altered DNA methylation patterns. Singleplex MethyLight analysis revealed significant PMR differences for MAGE-A1, LINE-1, and TGFβ2 between benzene-exposed workers and controls, highlighting the potential of DNA methylation profiling in occupational health risk assessment.