Abstract
The goal of early detection and screening is the diagnosis and treatment of cancer before it spreads beyond the organ of origin, perhaps even in its preinvasive state. Unfortunately, available early detection and screening techniques pick up many tumors at a relatively late stage in their natural history. As a result, decrements in mortality even with the best available detection modalities are likely to be modest. On the other hand, some early detection and screening techniques identify changes with a low probability of progression to life-threatening cancer, thereby resulting in unnecessary diagnosis and overtreatment. New technologies coming from the field of molecular and cellular biology are able to identify genetic as well as antigenic changes during the early stages of malignant progression. Some of these changes show promise as biomarkers for preneoplastic development or for malignant transformation.
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American Cancer Society. (1999) Cancer Facts & Figures—1999, American Cancer Society, Washington, DC.
Munger, K. (2002) The role of human papillomaviruses in human cancers. Front. Biosci. 7, d641–d649
Reeves, W. C, Rawls, W. E., and Brinton, L. A. (1989) Epidemiology of genital papillomaviruses and cervical cancer. Rev. Infect. Dis. 11, 426–439.
Muñoz, N., Bosch, F. X., de Sanjosé, S., et al. (1992) The causal link between human papillomavirus and invasive cervical cancer: a population-based case-control study in Columbia and Spain. Int. J. Cancer 52, 743–749.
Schiffman, M. H., Bauer, H. M., Hoover, R. N., et al. (1993) Epidemiologic evidence showing that human papillomavirus infection causes most cervical intraepithelial neoplasia. J. Natl. Cancer Inst. 85, 958–964.
Eluf-Neto, J., Booth, M., Muñoz, N., et al. (1994) Human papillomavirus and invasive cervical cancer in Brazil. Br. J. Cancer 69, 114–119.
Dyson, N., Howley, P. M., Münger, K., et al. (1989) The human papillomavirus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product. Science 243, 934–947.
Scheffner, M., Werness, B. A., Huibregtse, J. M., et al. (1990) The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell 63, 1129–1136.
Werness, B. A., Levine, A. J., and Howley, P. M. (1990) Association of human papillomavirus types 16 and 18 E6 proteins with p53. Science 248, 76–79.
Gao, Q., Srinivasan, S., Boyer, S. N., et al. (1999) The E6 oncoproteins of high-risk papillomaviruses bind to a novel putative GAP protein, E6TP1, and target for degradation. Mol. Cell. Biol 19, 733–744.
Pirisi, L., Creek, K. E., Doniger, J., et al. (1988) Continuous cell lines with altered growth and differentiation properties originate after transfection of human keratino-cytes with human papillomavirus type 16 DNA. Carcinogenesis 9, 1573–1579.
Woodworth, C. D., Bowden, P. E., Doniger, J., et al. (1988) Characterization of normal human exocervical cell immortalized in vitro by papillomavirus types 16 and 18 DNA. Cancer Res. 48, 4620–4628.
Halbert, C. L., Demers, G. W., and Galloway, D. A. (1991) The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65, 473–478.
Griep, A. E., Herber, R., Jeon, S., et al. (1993) Tumorigenicity by human papillomavirus type 16 E6 and E7 in transgenic mice correlates with alterations in epithelial cell growth and differentiation. J. Virol. 67, 1373–1384.
Arbeit, J. M., Munger, K., Howley, P. M., et al. (1994) Progressive squamous epithelial neoplasia in K14-human papillomavirus type 16 transgenic mice. J. Virol. 68, 4358–4368.
Herber, R., Liem, A., Pitot, H., et al. (1996) Squamous epithelial hyperplasia and carcinoma in mice transgenic for the human papillomavirus type 16 E7 oncogene. J. Virol. 70, 1873–1881.
Stoler, M. H., Rhodes, C. R., Whitbeck, A., et al. (1992) Human papillomavirus type 16 and 18 gene expression in cervical neoplasia. Hum. Pathol. 23, 117–128.
Unger, E. R., Vernon, S. D., Lee, D. R., et al. (1998) Detection of human papillomavirus in archival tissues: comparison of in situ hybridization and polymerase chain reaction. J. Histochem. Cytochem. 46, 535.
Ferenczy, A. and Franco, E. (2002) Persistent human papillomavirus infection and cervical neoplasia [review]. Lancet Oncol. 3, 11–16.
Lorenzato, F. R., Singer, A., Ho, L., et al. (2002) Human papillomavirus detection for cervical cancer prevention with polymerase chain reaction in self-collected samples. Am. J. Obstet. Gynecol. 186, 962–968.
Dimulescu, I., Unger, E. R., Lee, D. R., et al. (1998) Characterization of RNA in cytologic samples preserved in a methanol based collection medium. Mol. Diagn. 3, 1–7.
Xi, L. F., Carter, J. J., Galloway, D. A., et al. (2002) Acquisition and natural history of human papillomavirus type 16 variant infection among a cohort of female university students. Cancer Epidemiol. Biomarkers Prev. 11, 343–351.
Ting, Y. and Manos M. (1990) Detection and typing of genital human papillomaviruses, in PCR Protocols: A Guide to Methods and Applications (Innis, M. A., Gelfand, D. H., Sninsky, J. J., and White, T. J., eds.), Academic, New York, pp. 356–367.
Gravitt, P. E., Peyton, C. L., Alessi, T. Q., et al. (2000) Improved amplification of genital human papillomaviruses. J. Clin. Microbiol. 38, 357–361.
de Roda Husman, A. M., Walboomers, J. M., Meijer, C. J., et al. (1994) Analysis of cytomorphologically abnormal cervical scrapes for the presence of 27 mucosotropic human papillomavirus genotypes, using polymerase chain reaction. Int. J. Cancer 56, 802–806.
Kleter, B., van Doorn, L. J., ter Schegget, J., et al. (1998) A novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses. Am. J. Pathol. 153, 1731–1739.
Tucker, R. A., Johnson, P. R., Reeves, W. C., et al. (1993) Using the polymerase chain reaction to genotype human papillomavirus DNAs in samples containing multiple HPVs may produce inaccurate results. J. Virol. Methods 43, 321–333.
Vernon, S. D., Unger, E. R., and Williams, D. (2000) Comparison of human papillomavirus detection and typing by cycle sequencing, line blot and hybrid capture. J. Clin. Microbiol. 38, 651–655.
Coutlee, F., Gravitt, P., Kornegay, J., et al. (2002) Use of PGMY primers in L1 consensus PCR improves detection of human papillomavirus DNA in genital samples. J. Clin. Microbiol. 40, 902–907.
Jacobs, M. V., de Roda Husman, A. M., van den Brule, A. J., et al. (1995) Group-specific differentiation between high-and low-risk human papillomavirus genotypes by general primer-mediated PCR and two cocktails of oligonucleotide probes. J. Clin. Microbiol. 33, 901–905.
van den Brule, A. J., Pol, R., Fransen-Daalmeijer, N., et al. (2002) GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high-throughput identification of human papillomavirus genotypes. J. Clin. Microbiol. 40, 779–787.
Kleter, B., van Doorn, L. J., Schrauwen, L., et al. (1999) Development and clinical evaluation of a highly sensitive PCR-reverse hybridization line probe assay for detection and identification of anogenital human papillomavirus. J. Clin. Microbiol. 37, 2508–2517.
Gravitt, P. E., Peyton, C. L., Apple, R. J., et al. (1998) Genotyping of 27 human papillomavirus types by using L1 consensus PCR products by a single-hybridization, reverse line blot detection method. J. Clin. Microbiol. 36, 3020–3027.
van Doorn, L. J., Quint, W., Kleter, B., et al. (2002) Genotyping of human papillomavirus in liquid cytology cervical specimens by the PGMY line blot assay and the SPF(10) line probe assay. J. Clin. Microbiol. 40, 979–983.
Kerstens, H. M. J., Poddighe, P. J., and Hanselaar, A. G. J. M. (1995) A novel in situ hybridization method based on the deposition of biotinylated tyramide. J. Histochem. Cytochem. 43, 347.
Plummer, T. B., Sperry, A. C., Xu, H. S., et al. (1998) In situ hybridization detection of low copy nucleic acid sequences using catalyzed reporter deposition and its usefulness in clinical human papillomavirus typing. Diagn. Mol. Pathol. 7, 76.
Sano, T., Hikino, T., Niwa, Y., et al. (1998) In situ hybridization with biotinylated tyramide amplification: detection of human papillomavirus DNA in cervical neo-plastic lesions. Mod. Pathol. 11, 19.
Cheung, A. L. M., Graf, A.-H., Hauser-Kronberger, et al. (1999) Detection of human papillomavirus in cervical carcinoma: comparison of peroxidase, nanogold, and catalyzed reported deposition (CARD)-nanogold in situ hybridization. Mod. Pathol. 12, 689.
Unger, E. R., Hammer, M. L., and Chenggis, M. L. (1991) Comparison of 35S and biotin as labels for in situ hybridization: use of an HPV model system. J. Histochem. Cytochem. 39, 145.
Unger, E. R., Vernon, S. D., Hewan-Lowe, K. O., et al. (1999) An unusual cervical carcinoma showing exception to epitheliotropism of human papillomavirus. Hum. Pathol. 30, 483.
Unger, E. R., Vernon, S. D., Lee, D. R., et al. (1997) Human papillomavirus type in anal epithelial lesions is influenced by human immunodeficiency virus. Arch. Pathol. Lab. Med. 121, 820.
Unger, E. R., Vernon, S. D., Thoms, W. W., et al. (1995) Human papillomavirus and disease-free survival in FIGO stage Ib cervical cancer. J. Infect. Dis. 172, 1184–1190.
Southern, S. A., Graham, D. A., and Herrington, C. S. (1998) Discrimination of human papillomavirus types in low and high grade cervical squamous neoplasia. Diagn. Mol. Pathol. 7, 114.
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Vernon, S.D., Unger, E.R. (2004). Molecular Characterization of Human Papillomaviruses by PCR and In Situ Hybridization. In: Roulston, J.E., Bartlett, J.M.S. (eds) Molecular Diagnosis of Cancer. Methods in Molecular Medicine, vol 97. Humana Press. https://doi.org/10.1385/1-59259-760-2:159
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DOI: https://doi.org/10.1385/1-59259-760-2:159
Publisher Name: Humana Press
Print ISBN: 978-1-58829-160-8
Online ISBN: 978-1-59259-760-4
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