Methylation status of the E2 binding sites of HPV16 in cervical lesions determined with the Luminex® xMAP™ system

Snellenberg, Suzanne; Schütze, Denise M.; Claassen-Kramer, Debbie; Meijer, Chris J.L.M.; Snijders, Peter J.F.; Steenbergen, Renske D.M.

doi:10.1016/j.virol.2011.11.006

Cited by 34 publications

(40 citation statements)

References 62 publications

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“…Taken together, our results confirm data from previous studies reporting a higher frequency of HPV16 promoter methylation in cancer samples than in precancerous lesions or normal samples (28,(35)(36)(37). However, the proportion of cancer samples with methylated E2BSs varied from 20% to 90%, likely highlighting the variability in the methodology used in the different studies (44).…”

Section: Hpv16 E2bs Hrm-pcr Is a Reliable Methods For Methylation Assesupporting

confidence: 88%

“…We propose that HPV16 E2BS HRM-PCR alone is sufficient to evaluate the methylation status of HPV16 E2BS1, E2BS2, and Sp1BS from cervical smears. Pyrosequencing analysis also showed that the five CpGs always displayed similar methylation levels, indicating no position effect, as was previously described (37,45). This suggests a concerted regulation of DNA methylation at the E2BS1, E2BS2, and Sp1BS CpG sites.…”

Section: Hpv16 E2bs Hrm-pcr Is a Reliable Methods For Methylation Assesupporting

confidence: 81%

“…Some studies reported decreased E2BS methylation during lesion progression (30)(31)(32) or a trend of a lower risk of incident precancerous and cancerous lesions for patients with high E2BS methylation (33,34). On the contrary, methylation of the five CpGs located in the E2BS1 and E2BS2 and Sp1-binding site (Sp1BS) has frequently been associated with cervical cancer (28,29,(35)(36)(37)(38), and a gradual increase in methylation during lesion progression toward cancer has been reported in some studies (29,36,37,39). Nevertheless, most of these studies did not include the full spectrum of cervical disease progression from normal to cancer.…”

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confidence: 99%

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Methylation of Human Papillomavirus Type 16 CpG Sites at E2-Binding Site 1 (E2BS1), E2BS2, and the Sp1-Binding Site in Cervical Cancer Samples as Determined by High-Resolution Melting Analysis–PCR

et al. 2013

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Certain Alphapapillomavirus species classified as high-risk human papillomaviruses (HR-HPVs) have been recognized as the etiologic factors of invasive cervical carcinoma worldwide (1, 2). HR-HPV infections are frequent in young women but they are generally transient, with an estimated mean duration of incident infection of 16 months (3). Only persistent HR-HPV infections are associated with an increased risk of developing high-grade cervical lesions or cancer of the cervix (4-6). Persistent HR-HPV infections may be associated with microscopic abnormalities called low-grade squamous intraepithelial lesions (LSIL). These LSIL present a high rate of regression following HPV clearance but may progress toward high-grade squamous intraepithelial lesions (HSIL) if HR-HPV infection persists. Upon diagnosis, HSIL are treated mostly by excisional procedures to avoid the risk of progression toward invasive cancer (for a review, see reference 7).Among the HR-HPV types, HPV16 exhibits the highest persistence rate (8). Moreover, HPV16 is associated with an increased risk of developing precancerous and cancerous lesions (9) and is considered the most carcinogenic PV in humans (10). This is probably why HPV16 prevalence increases with the severity of cervical lesions (11), reaching 61% in invasive cervical carcinoma worldwide (12).HPV-associated carcinogenesis requires the continuous overexpression of the two main viral oncoproteins E7 and E6, which interact with many cellular proteins leading to the induction and the maintenance of a transformed phenotype in infected cells (for a review, see reference 13). E7 and E6 expression is regulated by the viral E2 protein through the early promoter (named p97 for HPV16) located in the long control region (LCR) of the PV genome. This promoter harbors four specific E2-binding sites (E2BSs) sharing the consensus sequence 5=-ACCG(N) 4 15). The three sites proximal to the TATA box have been shown to be involved in E2-mediated repression of the promoter activity, and E2BS1 and E2BS2 are probably the main sites to achieve this effect (for a review, see reference 16). From a mechanistic point of view, the binding of E2 to E2BS1 and E2BS2 induces a displacement of transcription activators, such as specificity protein 1 (Sp1) and TATA binding protein (TBP), from their binding sites, leading to a repression of E7 and E6 expression from the early promoter (17, 18). HR-HPV DNA integration has been described as a key step in the carcinogenesis process, since it generally results in the disruption of the E2 open reading frame (ORF) (19,20).Epigenetic alterations, and particularly aberrant DNA methylation, are frequently associated with tumor progression. Methylation of DNA mainly occurs on cytosines located in CpG dinucle-

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Section: Hpv16 E2bs Hrm-pcr Is a Reliable Methods For Methylation Assesupporting

confidence: 88%

Section: Hpv16 E2bs Hrm-pcr Is a Reliable Methods For Methylation Assesupporting

confidence: 81%

mentioning

confidence: 99%

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Methylation of Human Papillomavirus Type 16 CpG Sites at E2-Binding Site 1 (E2BS1), E2BS2, and the Sp1-Binding Site in Cervical Cancer Samples as Determined by High-Resolution Melting Analysis–PCR

et al. 2013

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“…Viral methylation, which generally leads to the silencing of viral genes, is one of many protective mechanisms exerted by human host cells to inhibit viral invasion [84][85][86][87][88][89][90][91]. The assessment of viral methylation has been described for various hrHPV types [90].…”

Section: Hpv Dna Methylation Analysis Methylation Analy-mentioning

confidence: 99%

Management of high-risk HPV-positive women for detection of cervical (pre)cancer

Luttmer

Strooper

Steenbergen

et al. 2016

Expert Review of Molecular Diagnostics

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Introduction: Primary HPV-testing has been shown to provide a superior detection of women at risk of cervical (pre)cancer compared to cytology-based screening. However, as most high-risk HPV infections are harmless, additional triage testing of HPV-positive women is necessary to identify those with cervical (pre)cancer. In this paper, we compare the performance, advantages and limitations of clinically relevant available triage strategies for HPV-positive women. Areas covered: Many different colposcopy triage strategies, comprising both microscopy-based and molecular (virus/host-related) markers, have been suggested: Pap cytology, p16/Ki-67 dual-stained cytology, HPV16/18 genotyping, viral DNA methylation and host cell DNA methylation. Literature search was limited to triage strategies that have achieved at least phase 2 of the five-phase framework for biomarker development and studies including large cohorts (≥100 hrHPV-positive women). Triage markers were stratified by sample type (cervical scrape, self-collected sample) and by study population (screening, non-attendee, referral). Expert commentary: At present, repeat Pap cytology and Pap cytology combined with HPV16/18 genotyping are the only triage strategies that have been robustly shown to be ready for implementation. Other strategies such as p16/Ki-67 dual-stained cytology and host cell DNA methylation analysis, with or without additional HPV16/18 genotyping, are attractive options for the near future. ARTICLE HISTORY

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“…DNA hypermethylation of the HPV genome is greater in invasive cervical carcinomas than in cervical dysplasias. Snellenberg et al showed that methylation of several HPV-16 E2 binding sites is significantly higher in invasive cervical cancer than in situ cervical lesions (7 ). The reasons for such observations are still under investigation.…”

Section: Hpv-18 E6mentioning

confidence: 99%

Genomics of Cervical Cancer and the Role of Human Papillomavirus Pathobiology

Uyar

Rader

2014

Clinical Chemistry

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Cervical cancer represents one of the most common malignancies in women worldwide and predominantly affects women of poor socioeconomic status. Persistent infection with high-risk human papillomavirus (HPV) 3 is an essential factor for development of cervical cancer. HPV infection is also associated with cancers of the vulva, vagina, anus, and oropharynx. Globally, an estimated 610 000 (4.8%) of the 12.7 million new cancer cases that occurred in 2008 could be attributed to HPV infection, and Ͼ500 000 of those cases represent cancers of the cervix alone (1 ). Within the last 2 decades, research has culminated in major advances in techniques for cervical cancer screening using HPV DNA and has led to a vaccine to prevent HPV infection. Yet, our strategies for the care and treatment of today's patients already infected with HPV need urgent attention. Women diagnosed with invasive and metastatic cervical cancer are in critical need of more-sophisticated prognostic markers, targeted therapeutic options, and more-accurate surveillance strategies. Viral Integration and Oncoprotein TargetsThere are about 120 HPV types, and types 16 and 18 are associated with 70% of the cases of cervical cancer worldwide. Many studies have shown that HPV-18 -associated cervical cancer is a strong independent prognostic factor for poorer disease-free survival for women undergoing surgery or radiation for early-stage invasive cervical cancer (2 ). The biology behind the poorer outcomes is unknown, but studies have suggested that sequence variation in HPV genes produces differences in their oncogenic potential. For example, HPV-18 E64 (E6 transforming protein) oncogene variants differentially regulate members of the Akt/P13K pathway and show increased expression of genes involved in cancer cell extravasation and metastasis (3, 4 ).HPV integration into the host genome is a critical step in cervical carcinogenesis and is found in almost all invasive cervical cancers. Integration frequently disrupts HPV E2 (E2 regulatory protein) gene expression, leading to increases in E6 and E7 viral oncoproteins, which in turn promote cellular immortalization and transformation. E6 and E7 (E7 transforming protein) gene overexpression contribute to marked genomic instability, accumulation of secondary mutations, and malignant transformation (5 ). In addition, the virus integrates into host genes and regulatory elements, which may cause structural alteration of the host genome and transcriptional deregulation of gene expression (6 ). The sites of integration occur frequently at common fragile sites in the genome, but these sites may be less random than originally appreciated. Further evaluation of HPV integration sites by the various HPV types could shed light on additional cancer-causing genes.The 8-kb HPV genomes have CpGs scattered throughout their genes. With advancing disease, these CpGs are increasingly methylated by the host cell's DNA methyltransferases. This methylation may alter the expression patterns of viral genes that are relevant to transforma...

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Methylation status of the E2 binding sites of HPV16 in cervical lesions determined with the Luminex® xMAP™ system

Cited by 34 publications

References 62 publications

Methylation of Human Papillomavirus Type 16 CpG Sites at E2-Binding Site 1 (E2BS1), E2BS2, and the Sp1-Binding Site in Cervical Cancer Samples as Determined by High-Resolution Melting Analysis–PCR

Methylation of Human Papillomavirus Type 16 CpG Sites at E2-Binding Site 1 (E2BS1), E2BS2, and the Sp1-Binding Site in Cervical Cancer Samples as Determined by High-Resolution Melting Analysis–PCR

Management of high-risk HPV-positive women for detection of cervical (pre)cancer

Genomics of Cervical Cancer and the Role of Human Papillomavirus Pathobiology

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