Human papillomavirus 16 L1 gene methylation as a potential biomarker for predicting anal intraepithelial neoplasia in men who have sex with men (MSM)

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.

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Potential biomarker of human papillomavirus 16 L1 methylation for prediction of anal intraepithelial neoplasia in men who have sex with men (MSM)

10.1101/2021.01.07.425707 ◽

2021 ◽

Author(s):

Arkom Chaiwongkot ◽

Parvapan Bhattarakosol ◽

Nittaya Phanuphak ◽

Tippawan Pankam

Keyword(s):

Human Papillomavirus ◽

Methylation Status ◽

Intraepithelial Neoplasia ◽

Anal Intraepithelial Neoplasia ◽

Gene Methylation ◽

Human Papillomavirus 16 ◽

Early Promoter ◽

Potential Biomarker ◽

Sex With Men ◽

Methylation Patterns

Quantitative measurement of human papillomavirus (HPV) 16 early promoter and L1 genes methylation were analyzed in anal cells collected from men who have sex with men (MSM) to determine the potential biomarker for screening of HPV related anal cancer. The methylation patterns of the HPV16 genes including early promoter (CpG 31, 37, 43, 52 and 58) and L1 gene (CpG 5600, 5606, 5609, 5615, 7136 and 7145) were analyzed in 178 anal samples with histology diagnosed as normal, anal intraepithelial neoplasia (AIN) 1, AIN2 and AIN3 by pyrosequencing assay. Low methylation levels of early promoter (<10%) and L1 genes (<20%) were found in all detected normal anal cells, while medium to high methylation (>20-60%) in early promoter was found 1.5% (1/67) and 5% (2/40) in AIN1 and AIN2-3 samples, respectively. Interestingly, slightly increased L1 gene methylation level (>20-60%) especially at HPV16 5'L1 regions CpGs 5600 and 5609 from normal to AIN3 were demonstrated. Moreover, negative correlation between high HPV16 L1 gene methylation at CpGs 5600, 5609, 5615 and 7145 and low percentage of CD4+ was found in AIN3 HIV positive cases. When compared methylation status of AIN2-3 to those of the normal/AIN1 lesion, the results indicated potential of using HPV16 L1 gene methylation as a biomarker for HPV related cancer screening.

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Prediction of Human Papillomavirus 16 E6 Gene Expression and Cervical Intraepithelial Neoplasia Progression by Methylation Status

International Journal of Gynecological Cancer ◽

10.1111/igc.0b013e31819d8a5c ◽

2009 ◽

Vol 19 (3) ◽

pp. 321-325 ◽

Cited By ~ 30

Author(s):

Pavla Hublarova ◽

Roman Hrstka ◽

Pavla Rotterova ◽

Leopold Rotter ◽

Marie Coupkova ◽

...

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Cervical Intraepithelial Neoplasia ◽

Methylation Status ◽

Gene Promoter ◽

Intraepithelial Neoplasia ◽

Hpv 16 ◽

Human Papillomavirus 16 ◽

E6 Gene ◽

Asymptomatic Women

Introduction:Human papillomavirus (HPV) infection represents the most important risk factor for the development of cervical intraepithelial neoplasia (CIN) and cervical cancer. We aimed to analyze the consequences of methylation of the E6 gene promoter in distinct stages of HPV-16-induced cellular transformation to assess its importance for disease progression.Methods:Human papillomavirus 16 was detected by sensitive polymerase chain reaction (PCR). Determination of E6 gene promoter methylation was analyzed by digestion with specific restriction endonuclease McrBC followed by PCR amplification. Expression of the E6 gene was determined by quantitative real-time PCR.Results:Of 103 cervical smears from asymptomatic women with no cytological and colposcopic abnormalities, 20.4% were HPV-16-positive. Human papillomavirus 16 was present in 44.4% of 18 patients with CIN I, in 62.2% of 143 patients with CIN II/III, and in 74.2% of 31 cervix carcinoma specimens. The incidence of HPV-16 in all lesions compared with asymptomatic women was statistically significant (P< 0.001, Pearsonχ2test). Methylation was detected in 81% (n = 21) of HPV-16-positive asymptomatic smears compared with 62.5% in CIN I (n = 8), 31.5% (n = 89) in CIN II/III, and 43.4% (n = 23) in carcinomas; a statistical significance between lesions and healthy women was found (P< 0.001, Pearsonχ2test). Expression of E6 mRNA correlated with methylation status (P= 0.010, Mann-WhitneyUtest).Conclusions:We conclude that methylation of the E6 gene promoter in HPV-16 genome is a predictive biomarker for cervical cancer progression by regulating the expression of the E6 oncogene.

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