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Immunocytochemical Analysis of the Cervical Pap Smear

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Cervical Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1249))

Abstract

Although immunostained cervical Pap smears are not yet FDA approved for clinical use, it is very likely that they will become widely employed in the near future to identify neoplastic squamous and iendocervical glandular cells when screening liquid-based cytological preparations (i.e., SurePath™ or ThinPrep™). The current problem with cytology complemented by high-risk human papillomavirus (HPV) testing is poor specificity. HPV testing provides superior sensitivity, but many women are infected with the virus, while very few have had persistent infections leading to carcinoma. Pathologists routinely use antibodies directed against the cyclin-dependent kinase inhibitor p16 (p16INK4a) or a combination of antibodies directed against topoisomerase-2-alpha and minichromosome maintenance protein-2 (as in ProEx™ C) to improve diagnostic precision and accuracy in cervical tissue biopsies. This chapter will describe the immunocytochemical methods used by our group to immunostain cervical Pap smears and provide significantly improved positive predictive value when screening for cervical cancer.

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References

  1. U.S. Cancer Statistics Working Group (2012) United States cancer statistics: 1999–2008 incidence and mortality web-based report. Department of Health and Human Services, Centers for Disease Control and Prevention, and National Cancer Institute, Atlanta, GA, http://www.cdc.gov/uscs

    Google Scholar 

  2. Saslow D, Solomon D, Lawson HW, Killackey M, Kulasingam SL, Cain J et al (2012) American cancer society, American society for colposcopy and cervical pathology, and American society for clinical pathology screening guidelines for the prevention and early detection of cervical cancer. Am J Clin Pathol 137(4):516–542

    Article  PubMed  Google Scholar 

  3. Sherman M, Lorincz A, Scott D, Wacholder S, Castle P, Glass A, Mielzynska-Lohnas I, Rush B, Schiffman M (2003) Baseline cytology, human Papillomavirus testing, and risk for cervical neoplasia: a 10-year cohort analysis. J Natl Cancer Inst 95(1):46–52

    Article  PubMed  Google Scholar 

  4. Pirog EC, Quint KD, Yantiss RK, Pirog EC, Quint KD, Yantiss RK (2010) P16/CDKN2A and Ki-67 enhance the detection of anal intraepithelial neoplasia and condyloma and correlate with human Papillomavirus detection by polymerase chain reaction. Am J Surg Pathol 34(10):1449–1455

    Article  PubMed  Google Scholar 

  5. Fakhry C, Rosenthal BT, Clark DP, Gillison ML (2011) Associations between oral HPV16 infection and cytopathology: evaluation of an oropharyngeal “Pap-test equivalent” in high-risk populations. Cancer Prev Res (Phila) 4(9): 1378–1384

    Article  Google Scholar 

  6. Kollar LM, Kahn JA (2008) Education about human Papillomavirus and human Papillomavirus vaccines in adolescents. Curr Opin Obstet Gynecol 20(5):479–483

    Article  PubMed  Google Scholar 

  7. Giuliano AR (2007) Human Papillomavirus vaccination in males. Gynecol Oncol 107 (2 Suppl 1):S24–S26

    Article  PubMed  CAS  Google Scholar 

  8. Moinian M, Andersch B (1982) Does cervix conization increase the risk of complications in subsequent pregnancies? Acta Obstet Gynecol Scand 61(2):101–103

    Article  PubMed  CAS  Google Scholar 

  9. Acharya G, Kjeldberg I, Hansen SM, Sørheim N, Jacobsen BK, Maltau JM (2005) Pregnancy outcome after loop electrosurgical excision procedure for the management of cervical intraepithelial neoplasia. Arch Gynecol Obstet 272(2):109–112

    Article  PubMed  Google Scholar 

  10. Suh-Burgmann EJ, Whall-Strojwas D, Chang Y, Hundley D, Goodman A (2000) Risk factors for cervical stenosis after loop electrocautery excision procedure. Obstet Gynecol 96 (5 Pt 1):657–660

    Article  PubMed  CAS  Google Scholar 

  11. Sano T, Oyama T, Kashiwabara K, Fukuda T, Nakajima T (1998) Expression status of p16 protein is associated with human Papillomavirus oncogenic potential in cervical and genital lesions. Am J Pathol 153(6):1741–1748

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  12. Keating J, Cviko A, Riethdorf S, Riethdorf L, Quade B, Sun D, Duensing S, Sheets E, Munger K, Crum C (2001) Ki-67, cyclin E, and p16INK4 are complimentary surrogate biomarkers for human Papillomavirus-related cervical neoplasia. Am J Surg Pathol 25(7): 884–891

    Article  PubMed  CAS  Google Scholar 

  13. Kruse AJ, Baak JP, de Bruin PC, Jiwa M, Snijders WP, Boodt PJ et al (2001) Ki-67 immunoquantitation in cervical intraepithelial neoplasia (CIN): A sensitive marker for grading. J Pathol 193(1):48–54

    Article  PubMed  CAS  Google Scholar 

  14. Ishimi Y, Okayasu I, Kato C, Kwon H, Kimura H, Yamada K, Song S (2003) Enhanced expression of Mcm proteins in cancer cells derived from uterine cervix. Eur J Biochem 270: 1089–1101

    Article  PubMed  CAS  Google Scholar 

  15. Santin A, Zhan F, Bignotti E, Siegel E, Cane S, Bellone S, Palmieri M, Anfossi S, Thomas M, Burnett A et al (2005) Gene expression profiles of primary HPV16- and HPV18-infected early stage cervical cancers and normal cervical epithelium: identification of novel candidate molecular markers for cervical cancer diagnosis and therapy. Virology 331(2):269–291

    Article  PubMed  CAS  Google Scholar 

  16. Shroyer K, Homer P, Heinz D, Singh M (2006) Validation of a novel immunocytochemical assay for topoisomerase II-alpha and minichromosome maintenance protein 2 expression in cervical cytology. Cancer 108(5): 324–330

    Article  PubMed  CAS  Google Scholar 

  17. Shi J, Liu H, Wilkerson M, Huang Y, Meschter S, Dupree W et al (2007) Evaluation of p16INK4a, minichromosome maintenance protein 2, DNA topoisomerase IIalpha, ProEX C, and p16INK4a/ProEX C in cervical squamous intraepithelial lesions. Hum Pathol 38(9):1335–1344

    Article  PubMed  CAS  Google Scholar 

  18. Pinto AP, Schlecht NF, Woo TY, Crum CP, Cibas ES (2008) Biomarker (ProEx C, p16(INK4A), and MiB-1) distinction of high-grade squamous intraepithelial lesion from its mimics. Mod Pathol 21(9):1067–1074

    Article  PubMed  CAS  Google Scholar 

  19. Shroyer K, Chivukula M, Ronnett B, Morgan T (2011) CINtec p16 cervical histology compendium & staining atlas. Roche/mtm labs

    Google Scholar 

  20. Guo M, Baruch A, Silva E, Jan Y, Ling E et al (2011) Efficacy of p16 and ProExC immunostaining in the detection of high-grade cervical intraepithelial neoplasia and cervical carcinoma. Am J Clin Pathol 135:212–220

    Article  PubMed  Google Scholar 

  21. Schmidt D, Bergeeron C, Denton K, Ridder R (2011) p16/Ki-67 dual-stain cytology in the triage of ASCUS and LSIL Papanicolaou cytology. Cancer Cytopathol 119:158–166

    Article  PubMed  Google Scholar 

  22. Singh M, Mockler D, Akalin A, Burke S, Shroyer L, Shroyer K (2012) Immunocytochemical colocalization of p16 and Ki-67 predicts CIN2/3 and AIS/adenocarcinoma. Cancer Cytopathol 120:26–34

    Article  PubMed  CAS  Google Scholar 

  23. Morgan T, Rozelle C, Schreiner A, Veyliotti A, Wachs, S, Pilliod R, Caughey A, Krum R, Berlin M Cost-benefit analysis of immunostaining SurePath Pap smears for p16 or ProEx C. Cancer Cytopathol (in preparation)

    Google Scholar 

  24. Zur HH (2002) Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer 2(5):342–350

    Article  Google Scholar 

  25. Herfs M, Yamamoto Y, Laury A, Wang X, Nucci MR, McLaughlin-Drubin ME, Münger K, Feldman S, McKeon FD, Xian W, Crum CP (2012) A discrete population of squamocolumnar junction cells implicated in the pathogenesis of cervical cancer. Proc Natl Acad Sci USA 109(26):10516–10521

    Article  PubMed  CAS  PubMed Central  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Pathology, Oregon Health & Science University, Mail Code L471, 3181 SW Sam Jackson, Portland, OR, 97239, USA

    Terry K. Morgan M.D., Ph.D.

  2. Department of Obstetrics and Gynecology, Oregon Health & Science University, Mail Code L471, 3181 SW Sam Jackson, Portland, OR, 97239, USA

    Terry K. Morgan M.D., Ph.D. & Michelle Berlin M.D., M.P.H.

Authors
  1. Terry K. Morgan M.D., Ph.D.
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  2. Michelle Berlin M.D., M.P.H.
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Corresponding author

Correspondence to Terry K. Morgan M.D., Ph.D. .

Editor information

Editors and Affiliations

  1. Touro University California, Vallejo, California, USA

    Daniel Keppler

  2. Touro University California, Vallejo, California, USA

    Athena W. Lin

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Morgan, T.K., Berlin, M. (2015). Immunocytochemical Analysis of the Cervical Pap Smear. In: Keppler, D., Lin, A. (eds) Cervical Cancer. Methods in Molecular Biology, vol 1249. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2013-6_15

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  • DOI: https://doi.org/10.1007/978-1-4939-2013-6_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2012-9

  • Online ISBN: 978-1-4939-2013-6

  • eBook Packages: Springer Protocols

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