DNA Aneuploidy and Integration of Human Papillomavirus Type 16 E6/E7 Oncogenes in Intraepithelial Neoplasia and Invasive Squamous Cell Carcinoma of the Cervix Uteri
Tóm tắt
Purpose: Increasingly deregulated expression of the E6-E7 oncogenes of high-risk human papillomaviruses (HR-HPVs) has been identified as the major transforming factor in the pathogenesis of cervical dysplasia and derived cancers. The expression of these genes in epithelial stem cells first results in chromosomal instability and induces chromosomal aneuploidy. It is speculated that this subsequently favors integration of HR-HPV genomes into cellular chromosomes. This in turn leads to expression of viral cellular fusion transcripts and further enhanced expression of the E6-E7 oncoproteins. Chromosomal instability and aneuploidization thus seems to precede and favor integration of HR-HPV genomes.
Experimental Design: To prove this sequential concept, we analyzed here the sequence of events of DNA aneuploidization and integration in a series of HPV-16-positive cervical dysplastic lesions and carcinomas. Eighty-five punch biopsies of HPV-16-positive cervical lesions (20 CIN1/2, 50 CIN3, and 15 CxCa) were analyzed for DNA ploidy by DNA flow cytometry and for integration of HPV E6/E7 oncogenes using the amplification of papillomavirus oncogene transcripts assay, a reverse transcription-PCR method to detect integrate-derived human papillomavirus oncogene transcripts.
Results: DNA aneuploidy and viral genome integration were both associated with increasing dysplasia (P < 0.001, χ2 test for trend). In addition, DNA aneuploidy was associated with increased viral integration (P < 0.01, Fisher’s exact test). Nineteen of 20 (95%) lesions with integrated viral genomes had aneuploid cell lines; however, only 19 of 32 (59%) lesions with aneuploid cell lines had integrated viral genomes.
Conclusions: These data support the hypothesis that aneuploidization precedes integration of HR-HPV genomes in the progression of cervical dysplasia. Accordingly, deregulated viral oncogene expression appears to result first in chromosomal instability and aneuploidization and is subsequently followed by integration of HR-HPV genomes in the affected cell clones.
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Tài liệu tham khảo
Rajagopalan H, Nowak MA, Vogelstein B, Lengauer C. The significance of unstable chromosomes in colorectal cancer. Nat Rev Cancer, 3: 695-701, 2003.
Anton M, Nenutil R, Rejthar A, Kopecny J, Ptackova B, Zaloudik J. DNA flow cytometry: a predictor of a high-risk group in cervical cancer. Cancer Detect Prev, 21: 242-6, 1997.
Bollmann R, Bollmann M, Henson DE, Bodo M. DNA cytometry confirms the utility of the Bethesda system for the classification of Papanicolaou smears. Cancer (Phila.), 93: 222-8, 2001.
Melsheimer P, Klaes R, von Knebel Doeberitz M, Bastert G. Prospective clinical study comparing DNA flow cytometry and HPV typing as predictive tests for persistence and progression of CIN I/II. Cytometry, 46: 166-71, 2001.
Bosch FX, Lorincz A, Munoz N, Meijer CJ, Shah KV. The causal relation between human papillomavirus and cervical cancer. J Clin Pathol (Lond.), 55: 244-65, 2002.
von Knebel Doeberitz M, Rittmuller C, Aengeneyndt F, Jansen-Durr P, Spitkovsky D. Reversible repression of papillomavirus oncogene expression in cervical carcinoma cells: consequences for the phenotype and E6–p53 and E7-pRB interactions. J Virol, 68: 2811-21, 1994.
Munger K, Basile JR, Duensing S, et al Biological activities and molecular targets of the human papillomavirus E7 oncoprotein. Oncogene, 20: 7888-98, 2001.
Mantovani F, Banks L. The human papillomavirus E6 protein and its contribution to malignant progression. Oncogene, 20: 7874-87, 2001.
Duensing S, Munger K. Centrosome abnormalities, genomic instability and carcinogenic progression. Biochim Biophys Acta, 1471: M81-8, 2001.
Duensing S, Munger K. The human papillomavirus type 16 E6 and E7 oncoproteins independently induce numerical and structural chromosome instability. Cancer Res, 62: 7075-82, 2002.
Ho GY, Bierman R, Beardsley L, Chang CJ, Burk RD. Natural history of cervicovaginal papillomavirus infection in young women. N Engl J Med, 338: 423-8, 1998.
Ostor AG. Natural history of cervical intraepithelial neoplasia: a critical review. Int J Gynecol Pathol, 12: 186-92, 1993.
zur Hausen H. Immortalization of human cells and their malignant conversion by high risk human papillomavirus genotypes. Semin Cancer Biol, 9: 405-11, 1999.
Duensing S, Lee LY, Duensing A, et al The human papillomavirus type 16 E6 and E7 oncoproteins cooperate to induce mitotic defects and genomic instability by uncoupling centrosome duplication from the cell division cycle. Proc Natl Acad Sci USA, 97: 10002-7, 2000.
Klaes R, Woerner SM, Ridder R, et al Detection of high-risk cervical intraepithelial neoplasia and cervical cancer by amplification of transcripts derived from integrated papillomavirus oncogenes. Cancer Res, 59: 6132-6, 1999.
Wentzensen N, Ridder R, Klaes R, Vinokurova S, Schaefer U, von Knebel Doeberitz M. Characterization of viral-cellular fusion transcripts in a large series of HPV16 and 18 positive anogenital lesions. Oncogene, 21: 419-26, 2002.
Romanczuk H, Howley PM. Disruption of either the E1 or the E2 regulatory gene of human papillomavirus type 16 increases viral immortalization capacity. Proc Natl Acad Sci USA, 89: 3159-63, 1992.
von Knebel Doeberitz M, Bauknecht T, Bartsch D, zur Hausen H. Influence of chromosomal integration on glucocorticoid-regulated transcription of growth-stimulating papillomavirus genes E6 and E7 in cervical carcinoma cells. Proc Natl Acad Sci USA, 88: 1411-5, 1991.
Jeon S, Lambert PF. Integration of human papillomavirus type 16 DNA into the human genome leads to increased stability of E6 and E7 mRNAs: implications for cervical carcinogenesis. Proc Natl Acad Sci USA, 92: 1654-8, 1995.
Jeon S, Allen-Hoffmann BL, Lambert PF. Integration of human papillomavirus type 16 into the human genome correlates with a selective growth advantage of cells. J Virol, 69: 2989-97, 1995.
Ziegert C, Wentzensen N, Vinokurova S, et al A comprehensive analysis of HPV integration loci in anogenital lesions combining transcript and genome-based amplification techniques. Oncogene, 22: 3977-84, 2003.
Duensing S, Duensing A, Crum CP, Munger K. Human papillomavirus type 16 E7 oncoprotein-induced abnormal centrosome synthesis is an early event in the evolving malignant phenotype. Cancer Res, 61: 2356-60, 2001.
Duensing S, Duensing A, Flores ER, Do A, Lambert PF, Munger K. Centrosome abnormalities and genomic instability by episomal expression of human papillomavirus type 16 in raft cultures of human keratinocytes. J Virol, 75: 7712-6, 2001.
Otto FJ. High-resolution analysis of nuclear DNA employing the fluorochrome DAPI. Methods Cell Biol, 41: 211-7, 1994.
Jacobs MV, Snijders PJ, van den Brule AJ, Helmerhorst TJ, Meijer CJ, Walboomers JM. 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. J Clin Microbiol, 35: 791-5, 1997.
Cullen AP, Reid R, Campion M, Lorincz AT. Analysis of the physical state of different human papillomavirus DNAs in intraepithelial and invasive cervical neoplasm. J Virol, 65: 606-12, 1991.