Aspekte der Molekularen Pathogenese des Zervixkarzinoms für neue Marker in der Krebsfrüherkennung und Diagnostik

 

 Buy Article Permissions and Reprints

Zusammenfassung

Die Mortalitätsraten des Zervixkarzinoms konnten durch die Einführung eines bevölkerungsweiten Screenings von Frauen mit Hilfe des sog. Pap-Tests drastisch gesenkt werden. Dennoch sind diese Untersuchungen mit einer hohen Rate falsch-positiver und falsch-negativer Testergebnisse behaftet, so daß technische Verbesserungen der bisher üblichen Screeninguntersuchungen dringend erforderlich sind. Sogenannte „high risk”-Typen der humanen Papillomviren (HR-HPVs) sind als die kausalen Agentien des Zervixkarzinoms erkannt worden. Der Nachweis dieser Virusinfektionen erlaubt es, prinzipiell gefährdete Frauen zu erfassen, dennoch kommen diese Infektionen bei etwa 5-30 % der weiblichen Bevölkerung vor und nur ein sehr kleiner Prozentsatz der Infizierten entwickelt eine klinisch relevante Läsion. Durch Aktivität zweier viraler Onkogene, E6 und E7, können einige wenige HR-HPV-infizierte Zellen in einem sehr langwierigen Prozeß nach und nach in Krebszellen umfunktioniert werden. In der Folge der Aktivität des viralen E7-Onkogens wird ein zelluläres Markerprotein (p16) in den dysplastischen Zellen deutlich verstärkt exprimiert. Mit Hilfe hochspezifischer Antikörper können daher unabhängig vom jeweiligen HR-HPV-Typ Zervixdysplasien und daraus abgeleitete Karzinome sowohl in der Histologie wie auch der Zytologie eindeutig nachgewiesen werden (CINtec-Verfahren). In weit fortgeschrittenen dysplastischen Läsionen kommt es oft zur Integration des HR-HPV-Genoms in das Genom der Wirtszelle. Dies ist mit der verstärkten Expression der viralen Onkogene verbundenen. Durch den Nachweis viraler mRNA-Moleküle, die von den integrierten viralen Genomen abgeschrieben werden, konnte ein Marker für besonders progressionsgefährdete Krebsvorstufen entwickelt werden (APOT-Verfahren). Auf der Basis dieser neuen Marker werden daher absehbar sehr sensitive, stabile, einfache und somit auch sehr kostengünstige Krebsvorsorgemaßnahmen entwickelt werden können.

Aspects of the molecular pathogenesis of cervical cancer to define new markers for cancer early detection and diagnosis

Summary

The mortality rates of cervical cancer could be drastically reduced by the implementation of population wide cytological screening test for women (Pap-Test). However, these screening tests are hampered by high rates of false positive and false negative results, pointing to the urgent demand for improved screening technologies. High risk human papillomaviruses were identified as the causal agents of cervical cancer. The detection of the viral infection allows to identify patients at risk, however, about 5-30 % of the normal female population harbours these viruses and only very few of these develop clinically relevant lesions. The activity of two viral oncogenes E6 and E7 initiates in a long term process neoplastic transformation in few of the HPV harbouring cells. As consequence of the expression of E7 a cellular marker protein (p16) is increasingly expressed in dysplastic cells. Monoclonal antibodies directed against p16 allow therefore to specifically identify dysplastic cells and derived invasive cancers in histological slides but also cytological smears (CINtec Assay). In advanced preneoplastic lesions HPV genomes are often integrated into cellular chromosomes. This leads to enhanced expression of the viral oncogenes. The detection of specific viral mRNA transcripts derived from integrated HPV genomes allows to identify preneoplastic lesions with a particularly high risk for progression to invasive cancers (APOT-assay). These findings will allow to establish highly sensitive, but specific and cost efficient new cancer early detection assays.

Literatur

• 1 Parkin D M, Pisani P, Ferlay J. Estimates of the worldwide incidence of 25 major cancers in 1990.  Int J Cancer. 1999;  80 827-841
  CrossrefPubMedGoogle Scholar
• 2 Anderson C M, Thornton J G. Screening for cervical cancer. Graphs may mislead [letter].  BMJ. 1994;  309 953-954
  PubMedGoogle Scholar
• 3 Nieminen P, Kallio M, Anttila A, Hakama M. Organised vs. spontaneous Pap-smear screening for cervical cancer: A case-control study.  Int J Cancer. 1999;  83 55-58
  CrossrefPubMedGoogle Scholar
• 4 Anttila A, Pukkala E, Soderman B, Kallio M, Nieminen P, Hakama M. Effect of organised screening on cervical cancer incidence and mortality in Finland, 1963-1995: recent increase in cervical cancer incidence.  Int J Cancer. 1999;  83 59-65
  CrossrefPubMedGoogle Scholar
• 5 Segnan N. Cervical cancer screening. Human benefits and human costs in the evaluation of screening programmes.  Eur J Cancer. 1994;  30 A 873-875
  CrossrefPubMedGoogle Scholar
• 6 Cuzick J, Meijer C J, Walboomers J M. Screening for cervical cancer [letter; comment].  Lancet. 1998;  351 1439-1440
  PubMedGoogle Scholar
• 7 Zuna R E. The Pap smear revisited. Controversies and recent developments.  Postgrad Med. 1984;  76 36-43, 46
  PubMedGoogle Scholar
• 8 O'Sullivan J P, Ismail S M, Barnes W S. et al . Interobserver variation in the diagnosis and grading of dyskaryosis in cervical smears: specialist cytopathologists compared with non-specialists.  J Clin Pathol. 1994;  47 515-518
  PubMedGoogle Scholar
• 9 O'Sullivan J P. Observer variation in gynaecological cytopathology.  Cytopathology. 1998;  9 6-14
  CrossrefPubMedGoogle Scholar
• 10 de Vet H C, Knipschild P G, Schouten H J, Koudstaal J, Kwee W S, Willebrand D, Sturmans F, Arends J W. Interobserver variation in histopathological grading of cervical dysplasia.  J Clin Epidemiol. 1990;  43 1395-1398
  CrossrefPubMedGoogle Scholar
• 11 de Vet H C, Knipschild P G, Schouten H J, Koudstaal J, Kwee W S, Willebrand D, Sturmans F, Arends J W. Sources of interobserver variation in histopathological grading of cervical dysplasia.  J Clin Epidemiol. 1992;  45 785-790
  CrossrefPubMedGoogle Scholar
• 12 Abulafia O, Sherer D M. Automated cervical cytology: meta-analyses of the performance of the AutoPap 300 QC System.  Obstet Gynecol Surv. 1999;  54 469-476
  CrossrefPubMedGoogle Scholar
• 13 Cenci M, Nagar C, Giovagnoli M R, Vecchione A. The PAPNET system for quality control of cervical smears: validation and limits.  Anticancer Res. 1997;  17 4731-4734
  PubMedGoogle Scholar
• 14 Halford J A, Wright R G, Ditchmen E J. Prospective study of PAPNET: review of 25 656 Pap smears negative on manual screening and rapid rescreening.  Cytopathology. 1999;  10 317-332
  CrossrefPubMedGoogle Scholar
• 15 Sherman M E, Schiffman M, Herrero R. et al . Performance of a semiautomated Papanicolaou smear screening system: results of a population-based study conducted in Guanacaste, Costa Rica [see comments].  Cancer. 1998;  84 273-280
  CrossrefPubMedGoogle Scholar
• 16 Östör A G. Natural history of cervical intraepithelial neoplasia: a critical review.  Int J Gynecol Pathol. 1993;  12 186-192
  PubMedGoogle Scholar
• 17 Gissmann L, Boshart M, Durst M, Ikenberg H, Wagner D, zur Hausen H. Presence of human papillomavirus in genital tumors.  J Invest Dermatol. 1984;  83 (Suppl) 26s-28s
  CrossrefPubMedGoogle Scholar
• 18 von Knebel Doeberitz M. Papillomaviruses in human disease: Part I. Pathogenesis and epidemiology of human papillomavirus infections.  Eur J Med. 1992;  1 415-423
  PubMedGoogle Scholar
• 19 von Knebel Doeberitz M, Rittmuller C, zur Hausen H, Durst M. Inhibition of tumorigenicity of cervical cancer cells in nude mice by HPV E6-E7 anti-sense RNA [letter].  Int J Cancer. 1992;  51 831-834
  PubMedGoogle Scholar
• 20 Walboomers J M, Jacobs M V, Manos M M. et al . Human papillomavirus is a necessary cause of invasive cervical cancer worldwide.  J Pathol. 1999;  189 12-19
  Google Scholar
• 21 Scheffner M, Romanczuk H, Munger K, Huibregtse J M, Mietz J A, Howley P M. Functions of human papillomavirus proteins.  Curr Top Microbiol Immunol. 1994;  186 83-99
  PubMedGoogle Scholar
• 22 Nakao Y, Yang X, Yokoyama M, Ferenczy A, Tang S C, Pater M M, Pater A. Induction of p16 during immortalization by HPV 16 and 18 and not during malignant transformation.  Br J Cancer. 1997;  75 1410-1416
  PubMedGoogle Scholar
• 23 Sano T, Oyama T, Kashiwabara K, Fukuda T, Nakajima T. Expression status of p16 protein is associated with human papillomavirus oncogenic potential in cervical and genital lesions.  Am J Pathol. 1998;  153 1741-1748
  PubMedGoogle Scholar
• 24 Klaes R, Friedrich T, Spitkovsky D. et al . Overexpression of p16^INK4a as Specific Marker for Dysplastic and Neoplastic epithelial cells of the Cervix Uteri.  Int J Cancer. (in press) 2001; 
  PubMedGoogle Scholar
• 25 Kiviat N B, Koutsky L A, Paavonen J A. et al . Prevalence of genital papillomavirus infection among women attending a college student health clinic or a sexually transmitted disease clinic.  J Infect Dis. 1989;  159 293-302
  PubMedGoogle Scholar
• 26 Ho G Y, Bierman R, Beardsley L, Chang C J, Burk R D. Natural history of cervicovaginal papillomavirus infection in young women.  N Engl J Med. 1998;  338 423-428
  CrossrefPubMedGoogle Scholar
• 27 Petry K U, Bohmer G, Iftner T, Flemming P, Stoll M, Schmidt R E. Human papillomavirus testing in primary screening for cervical cancer of human immunodeficiency virus-infected women, 1990-1998.  Gynecol Oncol. 1999;  75 427-431
  CrossrefPubMedGoogle Scholar
• 28 zur Hausen H. Papillomaviruses causing cancer: evasion from host-cell control in early events in carcinogenesis.  J Natl Cancer Inst. 2000;  92 690-698
  CrossrefPubMedGoogle Scholar
• 29 zur Hausen H. Immortalization of human cells and their malignant conversion by high risk human papillomavirus genotypes.  Semin Cancer Biol. 1999;  9 405-411
  CrossrefPubMedGoogle Scholar
• 30 Yokoyama M, Nakao Y, Yang X, Sun Q, Tsutsumi K, Pater A, Pater M M. Alterations in physical state and expression of human papillomavirus type 18 DNA following crisis and establishment of immortalized ectocervical cells.  Virus Res. 1995;  37 139-151
  CrossrefPubMedGoogle Scholar
• 31 Luft F, Gebert J, Schneider A, Melsheimer P, von Knebel Doeberitz M. Frequent allelic imbalance of tumor suppressor gene loci in cervical dysplasia.  Int J Gynecol Pathol. 1999;  18 374-380
  PubMedGoogle Scholar
• 32 von Knebel Doeberitz M, Oltersdorf T, Schwarz E, Gissmann L. Correlation of modified human papilloma virus early gene expression with altered growth properties in C4-1 cervical carcinoma cells.  Cancer Res. 1988;  48 3780-3786
  PubMedGoogle Scholar
• 33 von Knebel Doeberitz M, Gissmann L. Analysis of the biological role of human papilloma virus (HPV)-encoded transcripts in cervical carcinoma cells by antisense RNA.  Hamatol Bluttransfus. 1987;  31 377-379
  PubMedGoogle Scholar
• 34 von Knebel Doeberitz M, Rittmuller C, zur Hausen H, Durst M. Inhibition of tumorigenicity of cervical cancer cells in nude mice by HPV E6-E7 anti-sense RNA [letter].  Int J Cancer. 1992;  51 831-834
  PubMedGoogle Scholar
• 35 von Knebel Doeberitz M. Papillomaviruses in human disease: Part II. Molecular biology and immunology of papillomavirus infections and carcinogenesis [see comments].  Eur J Med. 1992;  1 485-491
  PubMedGoogle Scholar
• 36 von Knebel Doeberitz M, Bauknecht T, Bartsch D, zur Hau-sen 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. 1991;  88 1411-1415
  PubMedGoogle Scholar
• 37 Jeon S, Allen-Hoffmann B L, Lambert P F. Integration of human papillomavirus type 16 into the human genome correlates with a selective growth advantage of cells.  J Virol. 1995;  69 2989-2997
  PubMedGoogle Scholar
• 38 Jeon S, Lambert P F. 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. 1995;  92 1654-1658
  CrossrefPubMedGoogle Scholar
• 39 Reuter S, Bartelmann M. 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.  EMBO J. 1998;  17 215-222
  CrossrefPubMedGoogle Scholar
• 40 Klaes R, Woerner S M, 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. 1999;  59 6132-6136
  PubMedGoogle Scholar

Prof. Dr. med. Magnus von Knebel Doeberitz

Sektion für Molekulare Diagnostik und Therapie
Chirurgische Universitätsklinik Heidelberg

Im Neuenheimer Feld 110

D-69120 Heidelberg