High-risk Human Papillomavirus E7 Oncoprotein Detection in Cervical Squamous Cell Carcinoma

Ressler, Sigrun; Scheiden, R; Dreier, Kerstin; Laich, Andreas; Müller‐Holzner, Elisabeth; Pircher, Haymo; Morandell, Dieter; Stein, Ines; Viertler, Hans-Peter; Santer, Frédéric R.; Widschwendter, Andreas; Even, Jos; Jansen‐Dürr, Pidder; Capesius, C.; Zwerschke, Werner

doi:10.1158/1078-0432.ccr-07-1222

Cited by 35 publications

(29 citation statements)

References 23 publications

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“…However, they were not usually detected by direct antibody staining but rather by using surrogate markers for E7, such as mini-chromosome maintenance, proliferating cell nuclear antigen, and bromodeoxyuridine incorporation, as markers of DNA replication, or p16 INK4a , which has been found activated in cervical cancers as a result of functional inactivation of pRb by E7 (7). Indeed, this last marker has been widely used for staining of clinical biopsies to help in the diagnosis of CIN2 or CIN3, and it is now admitted that its detection perfectly matches the expression of E7 itself in the CIN lesions (8)(9)(10).…”

Section: Introductionmentioning

confidence: 99%

HPV16 E2 Is an Immediate Early Marker of Viral Infection, Preceding E7 Expression in Precursor Structures of Cervical Carcinoma

et al. 2010

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The viral E2 gene product plays a crucial role in the human papillomavirus (HPV) vegetative cycle by regulating both transcription and replication of the viral genome. E2 is a transcriptional repressor of the E6 and E7 viral oncogenes for HPV types 16 and 18, which are involved in cervical cancers. Using new polyclonal antibodies against the HPV16 E2 protein, we showed that E2 is expressed at various precursor stages of cervical carcinoma by immunohistochemistry on paraffin-embedded clinical samples. E2 was found to be highly expressed in the nuclei and cytoplasm of cells forming the intermediate and upper layers of cervical intraepithelial neoplasia (CIN). We could show that the expressions of E2 and p16INK4a (surrogate marker for oncogenic E7 expression) were exclusive in most of the cases, thus implying that E2 is not expressed together with high levels of E7. Moreover, we found that E2 is expressed in a subset of columnar cells adjacent to the CIN. We could show that expression of E2 is topologically distinct from the proliferation markers p63 and Ki67, whereas it coincides with the expression of cytokeratin K13, a marker of squamous cell differentiation. Expression of E2 also topologically coincides with episomal amplification of viral genomes in the upper layers of CIN1. These in vivo data thus validate previous assumptions of the crucial role of E2 in the early steps of HPV infection and of its negative link with expression of the viral E6 and E7 oncogenes. Cancer Res; 70(13); 5316-25. ©2010 AACR.

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Section: Introductionmentioning

confidence: 99%

HPV16 E2 Is an Immediate Early Marker of Viral Infection, Preceding E7 Expression in Precursor Structures of Cervical Carcinoma

et al. 2010

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“…Thus, there is a need for new technologies for cervical cancer screening. We have demonstrated in previous studies that high-risk HPV E7 proteins are regularly expressed in cervical SCCs and ACs and in their high-grade precursor lesions, suggesting that high-risk E7 oncoproteins are necessary for this cancer and may serve as new tumor markers (8,11,12,26). In the current communication, we addressed the question if E7 oncoproteins of the high-risk virus HPV18 can be detected in cervical smears by an innovative enzyme-linked immunosorbent assay (ELISA).…”

mentioning

confidence: 96%

“…Early studies have shown that immortalization by the E7 oncoprotein involves its ability to bind and thereby functionally inactivate cell cycle regulatory proteins such as the retinoblastoma tumor suppressor protein (9,11). Further work has demonstrated that E7 is located in both the cytoplasm and the nucleus (1,6,8,11,14,17,24,26,29,34,39,42). In keeping with these findings, it has been shown that HPV16 E7 is an integral part of many cellular protein complexes in the cytoplasm as well as in the nucleus and has multiple biochemical functions in the deregulation of pathways necessary for the oncogenic potential of the virus (reviewed in references 21 and 42).…”

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confidence: 99%

Detection of Human Papillomavirus Type 18 E7 Oncoprotein in Cervical Smears: a Feasibility Study

et al. 2012

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Persistent infections by high-risk human papillomaviruses (HPVs) are the main etiological factor for cervical cancer, and expression of HPV E7 oncoproteins was suggested to be a potential marker for tumor progression. The objective of this study was to generate new reagents for the detection of the HPV18 E7 oncoprotein in cervical smears. Rabbit monoclonal antibodies against recombinant E7 protein of HPV type 18 (HPV18) were generated and characterized using Western blotting, epitope mapping, indirect immunofluorescence, and immunohistochemistry. One clone specifically recognizing HPV18 E7 was used for the development of a sandwich enzyme-linked immunosorbent assay (ELISA). The assay was validated using recombinant E7 proteins of various HPV types and lysates from E7-positive cervical carcinoma cells. A total of 14 HPV18 DNA-positive cervical swab specimens and 24 HPV DNA-negative-control specimens were used for the determination of E7 protein levels by the newly established sandwich ELISA. On the basis of the average absorbance values obtained from all 24 negative controls, a cutoff above which a clinical sample can be judged E7 positive was established. Significant E7 signals 6-to 30-fold over background were found in 7 out of 14 abnormal HPV18 DNA-positive cervical smear specimens. This feasibility study demonstrates for the first time that HPV18 E7 oncoprotein can be detected in cervical smears. P ersistent infections by human papillomaviruses (HPVs) are the main etiologic factor for cervical cancer (41). Approximately 85% of cervical cancers are squamous cell carcinomas (SCCs) which arise from dividing keratinocytes in the squamous epithelium of the ectocervix and 15% are adenocarcinomas (ACs) which arise from glandular cells located in the endocervix (3, 25). About 40 HPV genotypes that can infect epithelial squamous and glandular cells in the cervical mucosa have been described. On the basis of epidemiological and biochemical data, only a subgroup of HPV types, referred to as high-risk HPVs, is associated with intraepithelial lesions that have a high potential for progression to invasive carcinoma. Infections by high-risk HPV genotypes have been detected in virtually all cervical cancers (37). At least 15 highrisk HPV types have been associated with these cancers. HPV type 16 (HPV16) and HPV18 are the most prevalent genotypes worldwide in SCCs as well as in ACs (23).A persistent infection with oncogenic HPVs is necessary for the development of cervical precancer and cancer (3, 37, 40, 41). Initial events of cervical carcinogenesis after viral infection by highrisk HPV types are specific changes that overcome the transcriptional control of viral gene expression in the infected keratinocytes (40). Inactivation of these cellular control functions permits deregulated transcription of the early viral genes E6 and E7. This event triggers reprogramming of cell proliferation, apoptosis, differentiation, metabolism, epigenetic reorganization, and genomic instability (21). These changes can support the integration ...

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“…New technologies for cervical cancer screening are urgently needed. Previous studies suggest that high-risk HPV E7 proteins are regularly expressed in cervical carcinoma and in their high-grade precursor lesions, suggesting that high-risk E7 oncoproteins are necessary for this cancer and may serve as new tumor biomarkers [10,[17][18][19][20]. In the present study, we developed and validated a new ELISA test capable for the simultaneous detection of the E7 proteins of HPV-16, HPV-18, and HPV-45.…”

Section: Introductionmentioning

confidence: 90%

A New Sandwich ELISA Test Simultaneously Detecting E7 Proteins of HPV-16, 18 and 45 in Cervical Smears

Metzger¹,

Pittl²,

Kaufmann³

et al. 2016

Clin Microbiol

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Persistent infections by high-risk papilloma viruses (HPV) are the main etiological factor for cervical cancer, and E7 oncoproteins were suggested as new markers for tumor progression. The objective of this study was to generate a new Enzyme-Linked Immunosorbent Assay (ELISA)-based detection system to monitor expression of the E7 proteins of the high-risk HPV (hrHPV) types HPV-16, HPV-18, and HPV-45 in cervical smears. Using a combination of rabbit monoclonal antibodies raised against E7 proteins of HPV-16 and HPV-18/HPV-45, respectively, a trivalent E7-ELISA was developed and validated, using recombinant E7 proteins of various HPV types and lysates from E7-positive cervical carcinoma cells. The amount of 0.5 picogram of E7 protein per well was determined as detection limit.The E7-ELISA was used to determine E7 protein levels in cervical smears obtained from a total of 67 women. E7 protein concentration was below the detection limit in all HPV-negative smears, and E7 protein concentrations above background were found in some HPV-positive cervical samples. Together the work described herein provides a new tool for the simultaneous detection of E7 proteins of the three most prevalent in cervical neoplasia hrHPV types.

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High-risk Human Papillomavirus E7 Oncoprotein Detection in Cervical Squamous Cell Carcinoma

Cited by 35 publications

References 23 publications

HPV16 E2 Is an Immediate Early Marker of Viral Infection, Preceding E7 Expression in Precursor Structures of Cervical Carcinoma

HPV16 E2 Is an Immediate Early Marker of Viral Infection, Preceding E7 Expression in Precursor Structures of Cervical Carcinoma

Detection of Human Papillomavirus Type 18 E7 Oncoprotein in Cervical Smears: a Feasibility Study

A New Sandwich ELISA Test Simultaneously Detecting E7 Proteins of HPV-16, 18 and 45 in Cervical Smears

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