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High-risk HPV infection modulates the promoter hypermethylation of APC, SFRP1, and PTEN in cervical cancer patients of North India

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Abstract

Persistent infection with oncogenic HPV and downregulation of tumor suppressor genes play an essential role in the development and progression of cervical cancer. The present study aimed to identify the promoter methylation status of APC, SFRP1, and PTEN which are important regulators of Wnt pathway and their association with high-risk HPV infection and gene expression. Methylation Specific PCR (MSP) and quantitative reverse transcription PCR (RT-qPCR) were used to detect methylation status and gene expression levels of APC, SFRP1, and PTEN in cervical cancer biopsies (110) and paired non-cancerous biopsies (28). APC promoter was methylated in 38%, SFRP1 in 95%, and PTEN in 55% of the cervical cancer biopsies. Our data showed a trend of a higher rate of methylation of the gene promoters in cervical cancer biopsies while; they were majorly un-methylated in non-cancerous biopsies. Corresponding to a higher rate of methylation in cancer biopsies, the gene expression levels of APC, SFRP1, and PTEN were reduced in cervical cancer samples in comparison to normal cervix tissues. Further, we observed that 97% cancer biopsies were HPV infected and high-risk type HPV16 and 18 infections were significantly positively associated with APC (p = 0.008 and p = 0.007), SFRP1 (p = 0.003 and p = 0.0067), and PTEN (p = 0.049 and p = 0.008) promoter methylation. APC, SFRP1, and PTEN promoter hyper-methylation is positively associated with high-risk HPV infection and inversely associated with gene expression. Our findings show that high-risk HPV infection promotes methylation of these genes and further promotes their silencing.

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Abbreviations

IHEC:

Institutional human ethical committee

qRT-PCR:

Quantitative real-time polymerase chain reaction

MSP:

Methylation specific PCR

HPV:

Human papillomavirus

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Acknowledgements

We would like to thanks Parul Ahuja, Geetanjali Singhal, Kiran Malik, Rajbala and all the staff of PGIMS, Rohtak for their assistance during sample collection. We also like to thanks all participants enrolled in the study.

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Authors and Affiliations

  1. Department of Genetics, Maharishi Dayanand University, Rohtak, 124001, Haryana, India

    Lokesh Kumari Kadian, Ritu Yadav, Shivkant Sharma & Chetna Yadav

  2. Departments of Obstetrics and Gynaecology, PGIMS, Rohtak, Haryana, India

    Smiti Nanda

  3. Department of Biosciences and Bioengineering, IIT Bombay, Mumbai, Maharashtra, India

    Gulshan Gulshan

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  1. Lokesh Kumari Kadian
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Contributions

Conceived of or designed study: LK; RY and SN. Performed research: LK; SS and CY. Analyzed data: LK and GG. Contributed new methods or models: LKand GG. Wrote the paper: LK; GG and RY.

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Correspondence to Ritu Yadav.

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The authors declare that they have no conflicts of interest.

Ethics approval

Ethical approval for conducting this study was obtained from Maharishi Dayanand University Institutional Human Ethical Committee (IHEC) with Number IHEC/2016/80-13.06.16.

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A written informed consent was obtained from all the enrolled cases of cervical cancer.

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Kadian, L.K., Yadav, R., Nanda, S. et al. High-risk HPV infection modulates the promoter hypermethylation of APC, SFRP1, and PTEN in cervical cancer patients of North India. Mol Biol Rep 47, 9725–9732 (2020). https://doi.org/10.1007/s11033-020-05960-z

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