Promoter methylation of APC, P16, and WT1 genes among high-risk HPV infected women of North-Eastern states of India.

BACKGROUND: Human papillomavirus (HPV) associated cervical cancer is the most prevalent gynaecological malignancy among women globally. Promoter hypermethylation of targeted genes may serve as a novel biomarker to monitor HPV infection dynamics which are likely to progress to HPV related cancers. METHODS: Cervical smear of representative 180 (90 persistent and 90 transient) HPV cases were included in the study from follow samples of eight North-Eastern states of India during June 2022 to September 2024. HPV positive DNA was detected using nested PCR and subsequently all nested PCR positive samples were subjected to High-risk HPV (HR-HPV) genotyping using real-time polymerase chain reaction (PCR) targeting 16 HR-HPV genotypes. Methylation status of Adenomatous Polyposis Coli (APC), P16, and WT1 genes was assessed by bisulphite modification of DNA followed by methylation-specific PCR. RESULTS: The study included 180 women, of average age 37.85±8.37 years. The highest HPV infection rate was found in women aged 31-40 years (38.88%). The HR-HPV types HPV 16, 18, 56, 58 and 67/33/52/58 were most persistent. Methylation of the APC gene was observed in 15% (14/90) of persistent HPV cases, while P16 methylation was found in 34% (31/90) of persistent and 28% (26/90) of transient cases. WT1 methylation was higher in transient (50%, 41/90) than persistent (36%, 33/90) HPV cases. No significant methylation differences were observed across HPV types, although HPV HPV 67/33/52/58 showed higher APC and P16 methylation than unmethylated cases. CONCLUSIONS: The study highlights a notable role of APC, P16 and WT1 gene methylation in HPV persistent and clearance, which may provide crucial insights into HPV-associated cancer progression and identify potential biomarkers for early detection.