Association between human papillomavirus and Epstein - Barr virus DNA and gene promoter methylation of RB1 and CDH1 in the cervical lesions: a transversal study

Abstract

BackgroundHuman papillomavirus (HPV) inactivates the retinoblastoma 1 (RB1) gene by promoter methylation and reduces cellular E-cadherin expression by overexpression of DNA methyltransferase 1 (DNMT1). The Epstein-Barr virus (EBV) is an oncogenic virus that may be related to cervical carcinogenesis. In gastric cancer, it has been demonstrated that E-cadherin gene (CDH1) hypermethylation is associated with DNMT1 overexpression by EBV infection. Our aim was to analyze the gene promoter methylation frequency of RB1 and CDH1 and verify the association between that methylation frequency and HPV and EBV infection in cervical lesions.MethodsSixty-five samples were obtained from cervical specimens: 15 normal cervices, 17 low-grade squamous intraepithelial lesions (LSIL), 15 high-grade squamous intraepithelial lesions (HSIL), and 18 cervical cancers. HPV and EBV DNA testing was performed by PCR, and the methylation status was verified by MSP.ResultsHPV frequency was associated with cervical cancer cases (p = 0.005) but not EBV frequency (p = 0.732). Viral co-infection showed a statistically significant correlation with cancer (p = 0.027). No viral infection was detected in 33.3% (5/15) of controls. RB1 methylated status was associated with cancer (p = 0.009) and HPV infection (p = 0.042). CDH1 methylation was not associated with cancer (p = 0.181). Controls and LSIL samples did not show simultaneous methylation, while both genes were methylated in 27.8% (5/18) of cancer samples. In the presence of EBV, CDH1 methylation was present in 27.8% (5/18) of cancer samples. Only cancer cases presented RB1 promoter methylation in the presence of HPV and EBV (33.3%).ConclusionsThe methylation status of both genes increased with disease progression. With EBV, RB1 methylation was a tumor-associated event because only the cancer group presented methylated RB1 with HPV infection. HPV infection was shown to be significantly correlated with cancer conditions. The global methylation frequency was higher when HPV was present, showing its epigenetic role in cervical carcinogenesis. Nevertheless, EBV seems to be a cofactor and needs to be further investigated.Virtual SlidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1159157579149317.

Putative biomarkers for cervical cancer: SNVs, methylation and expression profiles

“…The hypermethylation of the RB1 promoter was studied in 50 samples from patients with prior lesions in the cervix and 15 cervix samples from patients with normal cytology and colposcopy. The results demonstrated an increasing degree of methylation with the severity of the lesion (p = 0.009) as well as an association with HPV infection (p = 0.042) [28]. However, these results are still controversial [31].…”

The interaction between human papillomavirus and other viruses

“…In cervical cancer, PCR detection of EBV DNA or RNA is found in more than 60% of invasive squamous cell carcinoma (Aromseree et al, 2015; Khenchouche et al, 2013; McCormick et al, 2015; Santos et al, 2009; Sasagawa et al, 2000). A correlation between the detection of EBV and lesion severity has also been observed, with EBV detected in 8–14% of normal samples, 12.5% of low grade CIN lesions, and 21–38% of high grade CIN lesions (Khenchouche et al, 2013).…”

BLCAP arrests G1/S checkpoint and induces apoptosis through downregulation of pRb1 in HeLa cells

“…Rb protein has profound effect on multiple cellular processes and has been reported to regulate the expression of genes involved in cell cycle progression, differentiation, development, proliferation and apoptosis (27,28). Rb protein molecular weight is about 110 kDa, with localization in the nucleus.…”