Detection of Regulatory Protein p16/INK4A in the Dysplastic Cervical Squamous Cell Epithelium is a Diagnostic Tool for Carcinoma Prevention

Česká verze

Authors: J. Rajčáni ¹; M. Adamkov ²; J. Hybenová; E. Moráveková; Ľ. Lauko; D. Felcanová; M. Benčat
Authors place of work: Laboratórium patologickej anatómie, Alpha Medical a. s., Martin ; Virologický ústav SAV, Bratislava, emeritný vedecký pracovník ¹; Ústav histológie a embryológie Jesseniovej LFUK, Martin ²
Published in the journal: Čes.-slov. Patol., 45, 2009, No. 4, p. 101-107
Category: Původní práce

Summary

Parallel sections from 423 randomly selected blocks representing biopsies of 178 women with the diagnosis of cervical dysplasia and/or erosion were stained for p16 polypeptide. The p16/INK4A (inhibitory kinase 4) protein is a cellular division regulator, expression of which increases in the presence of oncoprotein E7, encoded by human papillomavirus (HPV). Expression of p16 protein was seen in the nuclei and cytoplasm of dysplastic squamous epithelium cells as well as in carcinoma cells. In 16.6% of erosion cases, the p16 antigen was present in the basal and suprabasal layer of the surrounding squamous epithelium revealing features of CIN I/LSIL. In CIN I/LSIL as classified by HE staining, the p16 antigen was found in 65 out of 80 (81%) cases. The p16 protein was typically seen in dysplastic basal and suprabasal cells encompassing a confluent layer in the lowest third segment of stratified epithelium. In CIN II and CIN III grouped as HSIL, the positive rate of p16 antigen presence was 95% (in 45 cases out of 47) and/or 100% (in each of 27 cases), respectively. The typical sign of p16 antigen distribution in HSIL was its staining over two thirds and/or throughout the whole dysplastic epithelium. Extensive staining for p16 antigen was registered within nuclei as well as cytoplasm of neoplastic cells in all 6 cervical squamous cell carcinomas, which were examined in many sections when being used as positive controls. Based on our experience, we consider the p16 antigen staining a helpful tool indicating dysplastic cells and estimating their extent.

Key words:
human papillomavirus – cervical epithelium – CIN/SIL – p16 protein – immunohistochemical staining

Zdroje

1. Bean S.M., Eltoum I., Whitlow L. et al.: Immunohistochemical expression of p16 and Ki-67 correlates with degree of anal intraepithelial neoplasia. Am J Surg Pathol 31, 2007, 555–561.

2. Cullen A., Reid R., Campion M. et al.: Analysis of the physical state of different human papillomavirus DNAs in epithelial and invasive cervical neoplasm. J Virol 65, 1991, s. 606–612.

3. Dourbar J.: The papillomavirus life cycle. J clin Virol. 32 (suppl), 2005, S7-S15.

4. Flores E.R., Lambert P.F.: Evidence for switch in the mode oh human papillomavirus DNA replication during the viral life cycle. J Virol 71, 1997, s. 7167–7179.

5. Dray M., Russel P., Dalrymple C. et al.: p16/INK4a as a complementarz marker of high grade intraepithelial lesions of the uterine cervix. I. Experience with squamous lesions in 189 consecutive cervical biopsies. Pathology 37, 2005, s. 112–124.

6. Dyson N., Howley, P.M., Munger, K. et al.: The human papillomavirus 16 E7 oncoprotein is able to bind the retinoblastoma gene product. Science 243, 1989, s. 934–937.

7. Golijow C.D., Abba, M.C., Mourón, S.A. et al.: C-myc gene amplification detected in preinvasive intraepithelial lesions. Int J Gynecol Cancer 11, 2001, 463–465.

8. Hopman A.H., Smedts F., Dignef W. et al.: Transition of high-grade cervical intraepithelial neoplasia in micro-invasive neoplasia to microinvasive carcinoma is characterized by integration of HPV 16/18 and numerical chromosome abnormalities. J Pathol 202, 204, s. 23–33.

9. Howley P.A., Lowy D.R.: Papillomaviruses, s. 2299–2354, in Knipe DM, Howley PM (ed), Fieldęs Virology, 5th edition, Walters Kluver/Lippincott, Wiliams and Wilkins, Philadelphia, 2007.

10. Ikeda K., Tate G., Suzuki T., Mitsuya T.: Coordinate expression of cytokeratin 8 and cytokeratin 17 immunohistochemical staining in cervical intraepithelial neoplasia and cervical squamous cell carcinoma: an immunohistochemical analysis and review of the literature. Gynecologic Oncol. 108, 598–602, 2008.

11. Jeon S., Lambert P.F.: Integration of HPV 16 DNA into the human genome leads to increased stability of E6/E7 mRNAs: implications for cervical carcinogenesis. Proc Natl Acad Sci USA 92, 1995, s. 1654–1658.

12. Keating J.T., Cviko A., Rietdorf S. et al.: K-67, cyclin E and p16/INK4 are complimentary surrogate biomarkers for human papillomavirus-related cervical neoplasia. Am J Surg Pathol 25, 2001, 884–891.

13. Kirnbauer R., Booy F., Cheng N. et al: Papillomavirus L1 major capsid protein self-assembles into virus particles that are high immunogenic. Proc Natl Acad Sci USA 89, 1992, s. 12180–12184.

14. Klaes, R., Friedrich, T, Spitkovski, D. et al.: Over expression of p16/INK4A as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri. Int. J Cancer 92, 276–284, 2001.

15. Koutsky I.A., Holmes K.K., Critchlow, C.W. et al.: A cohort study of the risk of cervical intraepithelial neoplasia grade 2 and 3 in relation to papillomavirus infection. New Engl J Med. 327, 1992, s. 1272–1278.

16. Kruse A.J., Baak J.P.A., deBruin P.C. et al.: Ki-67 immunoquantitation in cervical intraepithelial neoplasia (CIN): a sensitive marker for grading. J Pathol. 196, 2001, 48–54.

17. Kruse A.J., Baak J.P.A, Janssen E.A. et al.: Low and high risk CIN I and II lesions: prospective predictive value of grade, HPV and Ki-67. J Pathol 199, 2003, 462–470.

18. Kruse A.J., Skaland I., Janssen E.A. et al.: Quantitative molecular parameters to identify low and high risk early CIN lesions: role of markers of proliferative activity and differentiation and Rb availability. In J Gynecol Pathol 23, 2004, s.100–109.

19. Li Y., Nichols M.A., Shay J.W. et al.: Transcriptional repression of the D type cyclin dependent kinase p16 by retinoblastoma susceptibility gene product pRb. Cancer Res 54, 1994, s. 5816–5820.

20. Luff R.D.: The Bethesda System for reporting cervical vaginal cytologic diagnoses. Report of the 1991 Bethesda Workshop. Hum Pathol 23, 1992, s. 719–721.

21. NCI: National Cancer Institute Workshop: the 1988 Bethesda system for reporting cervical/vaginal cytologic diagnoses. J.A.M.A. 262, 1989, s. 931–934.

22. Meisels, A, Fortin, R.: Condylomatous lesions of the cervix and vagina. I. Cytologic patterns. Acta cytol. 20, 505–809, 1976.

23. Meisels, A Fortin, R, Roy, M: Condylomatous lesions of the cervix II. Cytologic, colposcopic and histopathologic study. Acta Cytol. 21, 379–390, 1977.

24. Meyers C., Laimins L.A.: In vitro systems for the study and propagation of human papillomaviruses. Curr Topics Microbiol Immunol 186, 1994, s. 199–215.

25. Mulvany, NJ, Allen, DG, Wilson, SM: Diagnostic utility of p16/INK4A: a reappraisal of its use in cervical biopsies. Pathology 40, 335–344, 2008.

26. Murphy N, Ring, M, Killalea AG, Uhlmann, V et al.: p16/INK4A as a marker for cervical dyskaryosis: CIN and cGIN in cervical biopsies and ThinPrep smears. J clin Pathol. 56, 56–63, 2003.

27. Murphy, N, Ring, M, Heffron, CCBB et al.: p16/INK4a, CDC6 and MCM5: predictive biomarkers in cervical preinvasive neoplasia and cervical cancer. J clin Pathol 58, 525–534, 2005.

28. Ondriáš F: Patológia cervixu maternice. Agentúra EURODOS, Bratislava, 2005.

29. O’Neill, CJ, McCluggage WG: p16 expression in the female genital tract and its value in diagnosis. Adv clin Pathol. 13, 8–15, 2006.

30. Phelps, W.C., Yee, C.L., Munger K. et al.: The human papillomavirus type 16 E7 gene encodes transactivation and transformation functions similar to those of adenovirus E1A. Cell 53, 1988, s. 539–647.

31. Purola, E., Savia, E: Cytology of gynecologic condyloma acuminatum. Acta Cytol. 21, 26–31, 1977.

32. Rajčáni, J.: Čeľaď Papillomaviridae. s. 324–330, in Rajčáni J., Čiampor F., Lekárska Virológia, Veda, Vyd. SAV, Bratislava, 2006.

33. Rodriguez A.C., Schiffman M., Herrero R. et al.: Rapid clearance of human papillomavirus and implications for clinical focus on persistent infections. J Natl Cancer Inst 100, 2008, s. 513–517.

34. Varnai A.D., Bollman M., Bankfalvi A. et al.: Predictive testing of cervical pre-cancer by detecting human papillomavirus E6/E7 mRNA in cervical cytologies up to high grade squamous intraepithelial lesions: diagnostic and prognostic implications. Oncol. Rep. 19, 2008, s. 457–465.

35. deVilliers E.M., Fauquet C.L., Brocker T., Bernard H.U., zurHausen H.: Classification of papillomaviruses. Virology 324Ł 2004, s. 17–27.

36. Waltz A.E., Lachago J., Bose S.: p16 and Ki-67 immunostaining is a useful adjunct in the assessment of biopsies for HPV associated anal intraepithelial neoplasia. Am J Surg Pathol 30, 795–801, 2006.

37. Werness B.A., Levine A.J., Howley P.M. et al.: Association of human papillomavirus types 16 and 18 E6 proteins with p53. Science 248, 1990, s. 76–79.

38. White A., Livanos F.M., Tlsty T.D.: Differential disruption of genomic integrity and cell cycle regulation in normal human fibroblasts by the HPV oncoproteins. Genes Dev 8, 1994, s. 666–677.

39. Wright T.C., Kurman R.J., Ferenczy A.: Precancerous lesions of the cervix, s. 253–321, in Kurman R.J. (ed), Blausteinęs Patology of the Female Genital Tract, 5th ed., Springer, New York/Berlin, 2001.

40. Yildiz, IZ, Usubutun A, Firet P et al.: Efficiency of immunohistochemical p16 expression and HPV typing in cervical squamous cell intraepithelial lesion grading and review of the literature. Pathol Res Pract 203, 445–449, 2007.

41. Zanotti S., Fiseler-Eckhoff A., Mannherz H.G.: Changes in the topological expression of markers of differentiation and apoptosis in defined stages of human cervical dysplázia and carcinoma. Gynecologic Oncology 89, 2003, 376–384.