Comparison of Human Papillomavirus Messenger RNA and DNA Detection: A Cross-sectional Study of 4,136 Women &amp;gt;30 Years of Age with a 2-Year Follow-up of High-Grade Squamous Intraepithelial Lesion

Molden, Tor; Kraus, Irene; Karlsen, Frank; Skomedal, Hanne; Nygård, Jan F.; Hagmar, Björn

doi:10.1158/1055-9965.epi-04-0410

Cited by 136 publications

(113 citation statements)

References 35 publications

(36 reference statements)

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“…The high levels of E6/E7 expression in CIN which we observed appear to be contradictory to the data obtained in several other studies (Falcinelli et al, 1993;Riethdorf et al, 2001;Sotlar et al, 2004;Molden et al, 2005;Andersson et al, 2006). The most probable explanation for this is the sampling strategy used (biopsies versus cervical scrapes).…”

Section: Figurecontrasting

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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Section: Figurecontrasting

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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“…No other species of HPV was detected by PCR amplification followed by DNA sequencing in these particular biopsies. This does not reflect the distribution of HPV in Norwegian tumour samples reported by others, although HPV16 is reported to be the most common high-risk HPV in Norwegian specimens, accounting for more than 50% of all cases (Kraus et al, 2004;Molden et al, 2005). Owing to the technical limitation of the PCR/sequencing assay, we neither can exclude the presence of other subspecies of HPV within our samples, nor can we exclude the presence of viral variants, sequence disruptions or very low HPV levels that escaped detection by the methods used.…”

Section: Discussioncontrasting

confidence: 57%

Gene expression reveals two distinct groups of anal carcinomas with clinical implications

et al. 2008

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Human papillomavirus (HPV) is a major aetiological agent in anal carcinomas. We here present a study of global gene expression using microarray hybridisation in a collection of anal carcinoma biopsies. Quantitative PCR was used to verify expression of selected genes. All biopsies contained integrated DNA of human papillomavirus subtype 16 (HPV16) and expressed HPV16 E7 mRNA. No other subspecies of HPV were detected in these 13 biopsies as assessed by PCR amplification and DNA sequencing. Unsupervised cluster analysis, based on global mRNA expression, divided the tumour biopsies into two distinct groups. Cluster analysis based on a number of high-risk HPV and/or E2F-regulated genes reproduced this biopsy grouping, suggesting that integrated HPV16 substantially influenced global gene expression in approximately half the biopsies studied. The levels of HPV16 E7 mRNA were significantly different between the two groups, but with considerable overlap. Thus, influence on global gene expression could not be absolutely ascribed to the expression level of HPV16. To investigate whether this distinction in gene expression had prognostic impact, we studied protein expression in an independent cohort of 55 anal carcinomas not included in the microarray study of two differentially expressed candidate genes, minichromosome maintenance complex component 7 (MCM7) and cyclin-dependent kinase inhibitor 2A (CDKN2A or p16). HPV status was assessed by in situ hybridisation. There was a significant association between in situ staining for HPV E7 mRNA and immunostaining for CDKN2A (p16) and MCM7 protein. CDKN2A (p16) mRNA was found significantly differentially expressed between the two tumour groups. However, cluster analysis on genes directly regulated by CDKN2A (p16) could not reproduce this split of biopsies into two groups, suggesting that the transcriptional regulatory activity of CDKN2A in these biopsies is inhibited. Furthermore, protein expression of CDKN2A (p16) could not be associated with survival. MCM7 is directly regulated by E2F and induced by HPV, and its mRNA was found differentially expressed between the two tumour groups. High level of MCM7 protein was found to be associated with both improved relapse-free survival (RFS, P ¼ 0.02) and cancer-specific survival (CSS, P ¼ 0.03) in anal cancer patients treated with radiation with or without additional chemotherapy.

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“…In this study, HPV DNA results were positive in about three times more samples than E6/E7 mRNA expression. Studying women under the age of 30 years, Molden et al (30) observed HPV E6/ E7 mRNA expression in approximately two out of three HPV DNA positive women, being similar to the findings observed for women older than 30 years (31) . In our study, 60% of the CIN2+ samples were positive for E6/ E7 mRNA.…”

Section: Discussionmentioning

confidence: 56%

“…Thus, HPV types are classified as high-risk (HR) or low-risk (LR) according to the risk of cancer in the infected hosts. The most common HR types are 16,18,31,33,45, which are detected in > 85% of the carcinomas worldwide (8) .…”

Section: Introductionmentioning

confidence: 99%

CerviCal Cytology, HPv DNa aND mrNa – a ComParative StuDy iN 162 PatieNtS

Martins¹

2012

J bras doenças sex transm

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Introduction: in contrast to the general improvement of the socioeconomic status of the Brazilian population, pathologies that are characteristic of poor health assistance persist. Among those, cervical cancer (CC) is emblematic; it still presents a persistently high incidence. Objective: to compare the performance of cervical cytology to HPV DNA and mRNA detection methods in 162 patients undergoing routine gynecological clinical practice. Methods: a total of 162 patients attended during routine gynecological examination in a private clinic in Florianópolis, Santa Catarina, Brazil, had cervical samples collected and processed for cytopathological and molecular tests, conventional PCR and NASBA. Positive samples positive for HPV DNA were submitted to Type-Specific PCR (TS-HPV PCR). Patients with altered smears were submitted to colposcopy and biopsy. Results: among the 162 samples, 19.8% (32/162) had altered smears, being 4/32 classified as ASC-H, 9/32 as ASC-US, 9/32 as LSIL and 10/32 as HSIL. Biopsies revealed nine cases of CIN I, nine CIN II and one CIN III, while seven were negative for cervical neoplasia. Overall, HPV DNA was detected in 38.3% (62/162) of the samples and HPV E6/E7 mRNA expression was found in 13.6% (22/162). Using TS-HPV PCR, HPV 16 was the most frequent type, found in 8% of the samples (5/62). Considering CIN2+ the gold-standard, cytology had 38.5% of specificity. Sensitivity and specificity of HPV-DNA PCR and NASBA were, respectively, 100% and 60%; 18.7% and 68.7%. Conclusion: mRNA E6/E7 expression was not a highly specific or sensitive marker for prevalent cervical disease while HPV DNA may be used for cervical cancer screening only in conjunction to more specific adjuvant tests.

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Comparison of Human Papillomavirus Messenger RNA and DNA Detection: A Cross-sectional Study of 4,136 Women >30 Years of Age with a 2-Year Follow-up of High-Grade Squamous Intraepithelial Lesion

Cited by 136 publications

References 35 publications

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

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

Gene expression reveals two distinct groups of anal carcinomas with clinical implications

CerviCal Cytology, HPv DNa aND mrNa – a ComParative StuDy iN 162 PatieNtS

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