Abstract
The human papillomavirus (HPV) 16 early promoter and L1 gene methylation were quantitatively measured using pyrosequencing assay in anal cells collected from men who have sex with men (MSM) to determine potential biomarkers for HPV-related anal cancer. The methylation patterns of HPV16 genes, including the early promoter (CpG 31, 37, 43, 52, and 58) and L1 genes (CpG 5600, 5606, 5609, 5615, 7136, and 7145), were analyzed in 178 anal samples. The samples were diagnosed as normal, anal intraepithelial neoplasia (AIN) 1, AIN2, and AIN3. Low methylation levels of the early promoter (< 10%) and L1 genes (< 20%) were found in all detected normal anal cells. In comparison, medium to high methylation (≥ 20–60%) in the early promoter was found in 1.5% (1/67) and 5% (2/40) of AIN1 and AIN2-3 samples, respectively. Interestingly, slightly increased L1 gene methylation levels (≥ 20–60%), especially at the HPV16 5’L1 regions CpGs 5600 and 5609, were demonstrated in AIN2-3 specimen. Moreover, a negative correlation between high HPV16 L1 gene methylation at CpGs 5600, 5609, 5615, and 7145 and a percentual CD4 count was found in AIN3 HIV positive cases. When comparing the methylation status of AIN2-3 to that of normal/AIN1 lesions, the results indicated the potential of using HPV16 L1 gene methylation as a biomarker for HPV-related cancer screening.
Highlights
Anal carcinoma is a rare disease found globally in men and women, with an incidence of < 1–2 cases per 100,000 [1]
Histology results were obtained from 123 samples classified as normal (n = 16), AIN1 (n = 67), AIN2 (n = 12), and AIN3 (n = 28)
A high methylation of the L1 gene was detected in both cervical cell lines regardless of the copy number of integrated HPV16
Summary
Anal carcinoma is a rare disease found globally in men and women, with an incidence of < 1–2 cases per 100,000 [1]. A high incidence of anal cancer was present in HIVinfected women and HIV-infected men who have sex with men (MSM), accounting for 30/ 100,000 and 131/100,000, respectively [2]. HPV16 L1 methylation for predicting anal intraepithelial neoplasia
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