Human papillomavirus type 16 (HPV 16) gene expression and DNA replication in cervical neoplasia: Analysis by in situ hybridization

Dürst, Matthias; Glitz, Dagmar; Schneider, Achim; Hausen, Harald zur

doi:10.1016/0042-6822(92)90688-l

Cited by 209 publications

(143 citation statements)

References 45 publications

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“…The reaction was heated at 95°C for 8 min. After 2 min at 50°C, a master mix containing 6 l of 10 ϫ labeling buffer, 5 l of dNTP (without dATP), 5 l of ␣-32 P dATP and 1 l of Klenow enzyme (Roche, Mannheim, Germany) was added. Probes were mixed thoroughly and incubated for 30 min at 50°C.…”

Section: Probe Synthesismentioning

confidence: 99%

“…Persistent viral infection in combination with strong, constitutive expression of the viral oncogenes E6 and E7 is a decisive step for malignant transformation. 5 The viral proteins deregulate cell proliferation and induce genetic instability, which promote the accumulation of mutations and aneuploidy. 6 These together with epigenetic alterations such as inactivation of genes by methylation or loss of imprinting have a profound effect on the gene expression profile of cells.…”

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confidence: 99%

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Detection of cancer‐related gene expression profiles in severe cervical neoplasia

Sopov

Magbagbeolu

et al. 2004

Intl Journal of Cancer

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The molecular signatures of 20 severe cervical intraepithelial neoplasia (CIN3) cases and 10 cervical squamous cell cancers were determined to define cancer-related gene expression profiles. RNAs extracted from microdissected tissues were amplified by SMART technology and used as probes for hybridization of commercially available cDNA array filters comprising 1,176 cancer-related genes. Ninetytwo differentially expressed genes were identified by comparison of pooled cDNA from CIN3 vs. cervical cancer. Heterogeneity in expression of this subset of genes was then analyzed for each biopsy using an algorithm for self-organizing maps. For several gene clusters, the expression pattern for CIN3 differed significantly from that of cancer. Moreover, hierarchical clustering revealed significant differences in distribution of CIN and cancer. Several CIN cases were more strongly related to cancer, suggesting that gene expression profiling may be useful for subdividing pathologically indistinguishable precancers into different biologic entities. This approach also provides a basis for the identification of putative prognostic markers and for targeted molecular therapy.

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Section: Probe Synthesismentioning

confidence: 99%

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confidence: 99%

Detection of cancer‐related gene expression profiles in severe cervical neoplasia

Sopov

Magbagbeolu

et al. 2004

Intl Journal of Cancer

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“…These mitotic disturbances are potentiated by HR-HPV E6. In situ hybridization analyses of precancers reveal at most only low levels of viral E6-E7 transcripts in basal and parabasal cells of low-grade lesions (CIN1) in comparison to high-grade lesions (CIN2/3) (Durst et al, 1992). This suggests that viral oncogene expression is suppressed.…”

Section: Introductionmentioning

confidence: 99%

Integration of the HPV16 genome does not invariably result in high levels of viral oncogene transcripts

et al. 2007

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Virus integration into the host genome is a characteristic step during cervical carcinogenesis. Experimental data provide evidence that integration could result in increased levels of oncogene (E6/E7) transcripts. This is the first study in which the level of viral transcripts is correlated to the physical state of the viral genome in cervical intraepithelial neoplasia (CIN) and cervical carcinomas (CxCa). Using the APOT-assay integrate-derived transcripts only were detected in 3/28 (11%) CIN and in 28/55 (51%) carcinomas, respectively. The remaining biopsies contained either episome-derived transcripts only or both mRNA species. SybrGreen real time reverse transcriptase-PCR assays were used to quantify viral gene expression for (i) all transcripts initiated from p97, (ii) full-length E6, (iii) E6*I and (iv) E5 transcripts. E6/E7 transcript levels showed a broad distribution but similar median values irrespective of histopathological grading and physical state of the viral genome. Biopsies with integrate-derived transcripts only generally lacked E5-specific mRNA. Our data do not support the hypothesis that HPV integration invariably results in high levels of oncogene transcripts. Instead, constitutive expression of oncogene transcripts rather than the level of expression appears to be decisive for transformation and the maintenance of the malignant phenotype.

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“…29 Two viral oncogenes, E6 and E7, are consistently expressed in HPV-associated cancer cells, and their expression is essentially required to initiate and maintain the malignant phenotype. 30,31 The oncogenic activity of these viral gene products rely on their interaction with specific cellular proteins (for reviews, see refs. 27 and 28).…”

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confidence: 99%

Overexpression of p16INK4A as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri

et al. 2001

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Cytological screening for cervical cancer or its precursors using Papanicolaou's smear test (Pap test) has been highly efficient to reduce the morbidity and mortality of cervical cancer. However, evaluation of the Pap test relies on subjective diagnostic parameters and is affected by a high rate of false-positive and false-negative results. More objective diagnostic parameters to identify truly dysplastic or neoplastic cells in cervical smears as well as in cervical biopsy samples would therefore avoid insecurity for many patients and the high screening costs associated with repeated testing. Cervical dysplasia is induced by persistent infections through highrisk types of human papillomaviruses (HPVs). Outgrowth of dysplastic lesions is triggered by increasing expression of two viral oncogenes, E6 and E7, which both interact with various cell cycle-regulating proteins. Among these is the retinoblastoma gene product pRB, which is inactivated by E7. pRB inhibits transcription of the cyclin-dependent kinase inhibitor gene p16 INK4a . Increasing expression of the viral oncogenes in dysplastic cervical cells might thus be reflected by increased expression of p16 INK4a . In line with this hypothesis, we observed marked overexpression of p16 INK4a in all cervical intraepithelial neoplasm (CIN) I lesions (n ‫؍‬ 47) except those associated with low-risk HPV types (n ‫؍‬ 7), all CIN II lesions (n ‫؍‬ 32), all CIN III lesions (n ‫؍‬ 60) and 58 of 60 invasive cervical cancers. In contrast, no detectable expression of p16 INK4a was observed in normal cervical epithelium (n ‫؍‬ 42), inflammatory lesions (n ‫؍‬ 48) and low-grade cervical lesions (CIN I) associated with low-risk HPV types (n ‫؍‬ 7). Dysplastic cells could also be identified in cervical smears using a specific p16 INK4a monoclonal antibody. These data demonstrate that p16 INK4a is a specific biomarker to identify dysplastic cervical epithelia in sections of cervical biopsy samples or cervical smears.

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Human papillomavirus type 16 (HPV 16) gene expression and DNA replication in cervical neoplasia: Analysis by in situ hybridization

Cited by 209 publications

References 45 publications

Detection of cancer‐related gene expression profiles in severe cervical neoplasia

Detection of cancer‐related gene expression profiles in severe cervical neoplasia

Integration of the HPV16 genome does not invariably result in high levels of viral oncogene transcripts

Overexpression of p16INK4A as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri

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