Type-dependent integration frequency of human papillomavirus genomes in cervical lesions.

Chromosomal integration of high-risk human papillomavirus (HR-HPV) genomes is believed to represent a significant event in the pathogenesis of cervical cancer associated with progression from preneoplastic lesions to invasive carcinomas. This hypothesis is based on experimental data suggesting that integration-dependent disruption of HR-HPV E2 gene functions is important to achieve neoplastic transformation and on clinical data gathered by analyzing lesions induced by human papillomavirus (HPV) 16 and 18 that revealed integrated viral genome copies in the vast majority of cervical cancer cells. However, a substantial fraction of cervical cancers is associated with other HR-HPV types for which virtually no data concerning their integration status have been reported so far. Here, we compared integration frequencies of the five most common oncogenic HPV types (HPV16, 18, 31, 33, and 45) in a series of 835 cervical samples using a specific mRNA-based PCR assay (Amplification of Papillomavirus Oncogene Transcripts). Most precancerous lesions displayed exclusively episomal viral genomes, whereas 62% of the carcinomas had integrated viral genomes. However, the frequency of integrated HR-HPV genomes showed marked differences for individual HR-HPV types. HPV16, 18, and 45 were found substantially more often in the integrated state compared with HPV types 31 and 33. The analysis of the median age of patients with high-grade precancerous lesions and invasive cancers suggests that precancers induced by HPV types 18, 16, and 45 progress to invasive cervical cancer in substantially less time compared with precancers induced by HPV types 31 and 33. These findings suggest that integration of oncogenic HPV genomes in cervical lesions is a consequence rather than the cause of chromosomal instability induced by deregulated HR-HPV E6-E7 oncogene expression. Distinct HR-HPV types apparently provoke chromosomal instability in their host cells to a different extent than is reflected by their integration frequencies in advanced lesions and the time required for CIN 3 lesions to progress to invasive cancer.

[1]  P. Howley,et al.  Repression of the Integrated Papillomavirus E6/E7 Promoter Is Required for Growth Suppression of Cervical Cancer Cells , 2000, Journal of Virology.

[2]  L. Banks,et al.  The Human Papillomavirus E6 protein and its contribution to malignant progression , 2001, Oncogene.

[3]  F. X. Bosch,et al.  Inverse relationship between human papillomavirus (HPV) type 16 early gene expression and cell differentiation in nude mouse epithelial cysts and tumors induced by HPV-positive human cell lines , 1991, Journal of virology.

[4]  David I. Smith,et al.  Acquisition of High-Level Chromosomal Instability Is Associated with Integration of Human Papillomavirus Type 16 in Cervical Keratinocytes , 2004, Cancer Research.

[5]  F. Radvanyi,et al.  MYC activation associated with the integration of HPV DNA at the MYC locus in genital tumors , 2006, Oncogene.

[6]  M. Mravunac,et al.  Transition of high‐grade cervical intraepithelial neoplasia to micro‐invasive carcinoma is characterized by integration of HPV 16/18 and numerical chromosome abnormalities , 2004, The Journal of pathology.

[7]  M. Stanley,et al.  Selection of cervical keratinocytes containing integrated HPV16 associates with episome loss and an endogenous antiviral response. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[8]  F. X. Bosch,et al.  Epidemiologic classification of human papillomavirus types associated with cervical cancer. , 2003, The New England journal of medicine.

[9]  M. Campion,et al.  Analysis of the physical state of different human papillomavirus DNAs in intraepithelial and invasive cervical neoplasm , 1991, Journal of virology.

[10]  Claude Fauquet,et al.  Classification of papillomaviruses. , 2004, Virology.

[11]  P. Howley,et al.  Mechanisms of Human Papillomavirus E2-Mediated Repression of Viral Oncogene Expression and Cervical Cancer Cell Growth Inhibition , 2000, Journal of Virology.

[12]  Steven Wolinsky,et al.  Human papillomavirus type 16 and 18 gene expression in cervical neoplasias. , 1992, Human pathology.

[13]  P. Howley,et al.  Suppression of cellular proliferation by the papillomavirus E2 protein , 1995, Journal of virology.

[14]  A. Venuti,et al.  HPV16 and HPV18 in genital tumors: Significantly different levels of viral integration and correlation to tumor invasiveness , 2002, Journal of medical virology.

[15]  David R. Scott,et al.  The elevated 10-year risk of cervical precancer and cancer in women with human papillomavirus (HPV) type 16 or 18 and the possible utility of type-specific HPV testing in clinical practice. , 2005, Journal of the National Cancer Institute.

[16]  Tom Freeman,et al.  Changes in cervical keratinocyte gene expression associated with integration of human papillomavirus 16. , 2002, Cancer research.

[17]  A. Vincent-Salomon,et al.  Human papillomavirus genotype as a major determinant of the course of cervical cancer. , 1998, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[18]  Magnus von Knebel Doeberitz,et al.  DNA Aneuploidy and Integration of Human Papillomavirus Type 16 E6/E7 Oncogenes in Intraepithelial Neoplasia and Invasive Squamous Cell Carcinoma of the Cervix Uteri , 2004, Clinical Cancer Research.

[19]  C. Peyton,et al.  Human Papillomavirus Type 16 Integration in Cervical Carcinoma In Situ and in Invasive Cervical Cancer , 2006, Journal of Clinical Microbiology.

[20]  S. Vinokurova,et al.  Characterization of viral-cellular fusion transcripts in a large series of HPV16 and 18 positive anogenital lesions , 2002, Oncogene.

[21]  C. Wheeler,et al.  Human papillomavirus type 16 infections and 2-year absolute risk of cervical precancer in women with equivocal or mild cytologic abnormalities. , 2005, Journal of the National Cancer Institute.

[22]  David I. Smith,et al.  Preferential integration of human papillomavirus type 18 near the c-myc locus in cervical carcinoma , 2003, Oncogene.

[23]  P. Lambert,et al.  Integration of human papillomavirus type 16 DNA into the human genome leads to increased stability of E6 and E7 mRNAs: implications for cervical carcinogenesis. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[24]  P. Lambert,et al.  Integration of human papillomavirus type 16 into the human genome correlates with a selective growth advantage of cells , 1995, Journal of virology.

[25]  Karl Münger,et al.  Biological activities and molecular targets of the human papillomavirus E7 oncoprotein , 2001, Oncogene.

[26]  M. Stanley,et al.  Properties of a non‐tumorigenic human cervical keratinocyte cell line , 1989, International journal of cancer.

[27]  P. Porter,et al.  Human papillomavirus and prognosis of invasive cervical cancer: a population-based study. , 2001, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[28]  P. Lazo,et al.  The molecular genetics of cervical carcinoma , 1999, British Journal of Cancer.

[29]  M. von Knebel Doeberitz,et al.  Detection of high-risk cervical intraepithelial neoplasia and cervical cancer by amplification of transcripts derived from integrated papillomavirus oncogenes. , 1999, Cancer research.

[30]  M. Bartelmann,et al.  APM‐1, a novel human gene, identified by aberrant co‐transcription with papillomavirus oncogenes in a cervical carcinoma cell line, encodes a BTB/POZ‐zinc finger protein with growth inhibitory activity , 1998, The EMBO journal.

[31]  T. Kessis,et al.  Expression of HPV16 E6 or E7 increases integration of foreign DNA. , 1996, Oncogene.

[32]  L Beardsley,et al.  Natural history of cervicovaginal papillomavirus infection in young women. , 1998, The New England journal of medicine.

[33]  B. Monk,et al.  Human papillomavirus type 18: association with poor prognosis in early stage cervical cancer. , 1997, Journal of the National Cancer Institute.

[34]  R. DeSalle,et al.  The carcinogenicity of human papillomavirus types reflects viral evolution. , 2005, Virology.

[35]  E. Blennow,et al.  Physical State of HPV16 and Chromosomal Mapping of the Integrated Form in Cervical Carcinomas , 2001, Diagnostic molecular pathology : the American journal of surgical pathology, part B.

[36]  S. Vinokurova,et al.  Systematic Review of Genomic Integration Sites of Human Papillomavirus Genomes in Epithelial Dysplasia and Invasive Cancer of the Female Lower Genital Tract , 2004, Cancer Research.

[37]  C. Meijer,et al.  A general primer GP5+/GP6(+)-mediated PCR-enzyme immunoassay method for rapid detection of 14 high-risk and 6 low-risk human papillomavirus genotypes in cervical scrapings , 1997, Journal of clinical microbiology.

[38]  M. von Knebel Doeberitz,et al.  Overexpression of p16INK4A as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri , 2001, International journal of cancer.

[39]  B. Monk,et al.  Early stage cervical cancers containing human papillomavirus type 18 DNA have more nodal metastasis and deeper stromal invasion. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[40]  J. D. Benson,et al.  Papillomavirus E2 induces senescence in HPV‐positive cells via pRB‐ and p21CIP‐dependent pathways , 2000, The EMBO journal.

[41]  M. von Knebel Doeberitz,et al.  Influence of chromosomal integration on glucocorticoid-regulated transcription of growth-stimulating papillomavirus genes E6 and E7 in cervical carcinoma cells. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Harald zur Hausen,et al.  Papillomaviruses Causing Cancer: Evasion From Host-Cell Control in Early Events in Carcinogenesis , 2000 .

[43]  F. Guijon,et al.  Quantification of HPV‐16 E6‐E7 transcription in cervical intraepithelial neoplasia by reverse transcriptase polymerase chain reaction , 1993, International journal of cancer.

[44]  M. Schiffman,et al.  Chapter 5: Updating the natural history of HPV and anogenital cancer. , 2006, Vaccine.

[45]  H. Skomedal,et al.  Presence of E6 and E7 mRNA from Human Papillomavirus Types 16, 18, 31, 33, and 45 in the Majority of Cervical Carcinomas , 2006, Journal of Clinical Microbiology.

[46]  Karl Münger,et al.  Human papillomavirus immortalization and transformation functions. , 2002, Virus research.

[47]  M. Frohman,et al.  Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[48]  K. Münger,et al.  Centrosome abnormalities and genomic instability induced by human papillomavirus oncoproteins. , 2003, Progress in cell cycle research.

[49]  M. Hoeckel,et al.  A comprehensive analysis of HPV integration loci in anogenital lesions combining transcript and genome-based amplification techniques , 2003, Oncogene.

[50]  M. Yaniv,et al.  Different mechanisms contribute to the E2-mediated transcriptional repression of human papillomavirus type 18 viral oncogenes , 1997, Journal of virology.