Promoter methylation of tumor suppressor genes induced by human papillomavirus in cervical cancer

Yanatatsaneejit,Pattamawadee; Chalertpet,Kanwalat; Sukbhattee,Juthamard; Nuchcharoen,Irin; Phumcharoen,Piyathida; Mutirangura,Apiwat

doi:10.3892/ol.2020.11625

Promoter methylation of tumor suppressor genes induced by human papillomavirus in cervical cancer

Authors:
- Pattamawadee Yanatatsaneejit
- Kanwalat Chalertpet
- Juthamard Sukbhattee
- Irin Nuchcharoen
- Piyathida Phumcharoen
- Apiwat Mutirangura
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Affiliations: Human Genetics Research Group, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, Inter‑Department of Biomedical Sciences, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand, Center of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Published online on: May 14, 2020 https://doi.org/10.3892/ol.2020.11625
Pages: 955-961
Copyright: © Yanatatsaneejit et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cervical cancer is the most fourth common cancer in women worldwide. The E6 and E7 high-risk human papillomavirus (HPV) types are the main cause of this cancer. Several studies have revealed that promoter methylation of tumor suppressor genes is induced by HPV E7. Recently, it was found that HPV16‑E7 and the DNA methyltransferase 1 complex could bind at the cyclin A1 (CCNA1) promoter, resulting in CCNA1 promoter methylation. Therefore, there is a need to study other tumor suppressor genes for which HPV may induce promoter methylation. The present study investigated whether HPV induced cell adhesion molecule 1 (CADM1) and death associated protein kinase 1 (DAPK1) promoter methylation. C33a (no HPV infection) and SiHa (HPV 16 infection) cell lines were used for methylation status and expression observation. It was found that CADM1 and DAPK1 promoter methylation, no expression of CADM1 and decreased expression of DAPK1, was presented in SiHa cells. While no promoter methylation of these two genes was observed in C33a cells, with positive expression of the genes. It was subsequently investigated whether E6 and/or E7 could induce promoter methylation and decrease the expression of these two genes. Methylation‑specific primer PCR and quantitative PCR were performed to elucidate the promoter methylation status and expression of CADM1 and DAPK1 in C33a cells transfected with HPV16 E6‑PCDNA3 or HPV16 E7‑PCDNA3.1 myc‑his, compared to empty vector‑transfected cells. The results showed that HPV E7 could induce CADM1 promoter methylation and decrease the gene expression in HPV E7 transfected C33a cells, while HPV E6 could induce DAPK1 promoter methylation and decrease its expression in C33a cells transfected with HPV E6. Finally, the mechanism by which HPV E7 induced CADM1 promoter methylation was observed by performing chromatin immunoprecipitation; the data showed that E7 induced CADM1 methylation by the same mechanism as that for CCNA1, by binding at the CADM1 promoter, resulting in the subsequent reduction of its expression in cervical cancer.

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