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Sec62/Ki67 and p16/Ki67 dual-staining immunocytochemistry in vulvar cytology for the identification of vulvar intraepithelial neoplasia and vulvar cancer: a pilot study

  • Ferenc Zoltan TakacsEmail author
  • Julia Caroline Radosa
  • Florian Bochen
  • Ingolf Juhasz-Böss
  • Erich-Franz Solomayer
  • Rainer M. Bohle
  • Georg-Peter Breitbach
  • Bernard Schick
  • Maximilian Linxweiler
General Gynecology
  • 39 Downloads

Abstract

Purpose

The aim of this study was to analyze the diagnostic performance of a newly established immunocytochemical dual-staining protocol for the simultaneous expression of SEC62 and Ki67 in vulvar liquid-based cytology specimens for the identification of vulvar intraepithelial neoplasia (VIN) and vulvar cancer. In addition, we investigated the p16/Ki67 dual stain, which has already been established in cervical cytology.

Materials and methods

For this pilot study, residual material from liquid-based cytology was collected retrospectively from 45 women. The presence of one or more double-immunoreactive cells was considered as a positive test result for Sec62/Ki67 and p16/Ki67 dual staining. The test results were correlated with the course of histology.

Results

All cases of VIN and vulvar cancer were Sec62/Ki67 and p16/Ki67 dual-stain positive, and normal and low-grade squamous intraepithelial lesions were all negative. The sensitivity of cytology for VIN + cases was 100% (22/22), whereas punch biopsy classified one case of vulvar carcinoma as inflammation. All cases with high-intensity (grades 3 and 4) Sec62 staining in Sec62/Ki67-positive cases were carcinomas.

Conclusions

The results of this study demonstrate that Sec62/Ki67 and p16 Ki67 dual-staining cytology could be a promising adjunctive diagnostic tool for VIN and squamous cell carcinoma, in addition to standard histology.

Keywords

Sec62 Ki67 Immunocytochemistry Vulvar cytology Vulvar intraepithelial neoplasia Vulvar cancer 

Notes

Acknowledgements

The authors would like to gratefully thank for the excellent work, motivation and enthusiasm of Mrs. Barbara Linxweiler and Mrs. Alice Kunz in the establishment of our dual-staining cytology method.

Author contributions

FZT: conceptualization, data curation, methodology, project administration, validation, writing the original draft, and review and editing. JCR: review and editing. FB: visualization, review and editing. IJ-B: review and editing, E-FS: review and editing, RMB: review and editing. G-PB: review and editing. BS: review and editing. ML: supervision, methodology, project administration, validation, review and editing.

Compliance with ethical standards

Conflict of interest

Ferenc Zoltan Takacs and Maximilian Linxweiler have a pending patent application. Patent applicant: University of Saarland. Names of inventors: Ferenc Zoltan Takacs, Maximilian Linxweiler. Application number: LU100824. All other authors declare no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study was approved by the Ethics Committee of Saarland (No: 183/17). Written informed consent was obtained from all patients prior to inclusion.

Informed consent

The study was approved by the Ethics Committee of Saarland (No: 183/17). Written informed consent was obtained from all patients prior to inclusion.

References

  1. 1.
    Siegel RL, Miller KD, Jemal A (2017) Cancer statistics, 2017. CA Cancer J Clin 67:7–30PubMedPubMedCentralCrossRefGoogle Scholar
  2. 2.
    Rakislova N, Clavero O, Alemany L et al (2017) Histological characteristics of HPV-associated and-independent squamous cell carcinomas of the vulva: a study of 1,594 cases. Int J Cancer 141(12):2517–2527PubMedCrossRefGoogle Scholar
  3. 3.
    Buttmann-Schweiger N, Klug SJ, Luyten A, Holleczek B, Heitz F, du Bois A, Kraywinkel K (2015) Incidence patterns and temporal trends of invasive nonmelanotic vulvar tumors in Germany 1999–2011. A population-based cancer registry analysis. PloS One 10(5):e0128073PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Crum CP, Herrington CS, McCluggage WG, Regauer S, Wilkinson EJ (2014) Chapter 9, tumours oh the vulva, epithelial tumours. In: Kurman RJ, Carcangiu ML, Herrington CS, Young RH (eds) WHO classification of tumours of female reproductive organs. IARC, LyonGoogle Scholar
  5. 5.
    Kang Y-J, Smith M, Barlow E, Coffey K, Hacker N, Canfell K (2017) Vulvar cancer in high-income countries: increasing burden of disease. Int J Cancer 141(12):2174–2186PubMedCrossRefGoogle Scholar
  6. 6.
    Edwards SK, Bates CM, Lewis F, Sethi G, Grover D (2015) 2014 UK national guideline on the management of vulval conditions. Int J STD AIDS 26(9):611–624PubMedCrossRefGoogle Scholar
  7. 7.
    Polterauer S, Dressler AC, Grimm C, Seebacher V, Tempfer C, Reinthaller A, Hefler L (2009) Accuracy of preoperative vulva biopsy and the outcome of surgery in vulvar intraepithelial neoplasia 2 and 3. Int J Gynecol Pathol 28(6):559–562PubMedCrossRefGoogle Scholar
  8. 8.
    Preti M, Bucchi L, Ghiringhello B, Privitera S, Frau V, Corvetto E, Benedetto C, Micheletti L (2017) Risk factors for unrecognized invasive carcinoma in patients with vulvar high-grade squamous intraepithelial lesion at vulvoscopy-directed biopsy. J Gynecol Oncol 28(4):e27PubMedPubMedCentralCrossRefGoogle Scholar
  9. 9.
    Jee KJ, Kim YT, Kim KR, Kim HS, Yan A, Knuutila S (2001) Loss in 3p and 4p and gain of 3q are concomitant aberrations in squamous cell carcinoma of the vulva. Mod Pathol 14(5):377–381PubMedCrossRefGoogle Scholar
  10. 10.
    Heselmeyer K, Macville M, Schröck E et al (1997) Advanced-stage cervical carcinomas are defined by a recurrent pattern of chromosomal aberrations revealing high genetic instability and a consistent gain of chromosome arm 3q. Genes Chromosomes Cancer 19(4):233–240PubMedCrossRefGoogle Scholar
  11. 11.
    Hopman A, Theelen W, Hommelberg P, Kamps M, Herrington C, Morrison L, Speel E-J, Smedts F, Ramaekers F (2006) Genomic integration of oncogenic HPV and gain of the human telomerase gene TERC at 3q26 are strongly associated events in the progression of uterine cervical dysplasia to invasive cancer. J Pathol 210(4):412–419PubMedCrossRefGoogle Scholar
  12. 12.
    Linxweiler M, Bochen F, Schick B et al (2016) Identification of SEC62 as a potential marker for 3q amplification and cellular migration in dysplastic cervical lesions. BMC Cancer 16(1):676PubMedPubMedCentralCrossRefGoogle Scholar
  13. 13.
    Hagerstrand D, Tong A, Schumacher SE, Ilic N, Shen RR, Cheung HW, Vazquez F, Shrestha Y, Kim SY, Giacomelli AO, Rosenbluh J, Schinzel AC, Spardy NA, Barbie DA, Mermel CH, Weir BA, Garraway LA, Tamayo P, Mesirov JP, Beroukhim R, Hahn WC (2013) Systematic interrogation of 3q26 identifies TLOC1 and SKIL as cancer drivers. Cancer Discov 3(9):1044–1057PubMedPubMedCentralCrossRefGoogle Scholar
  14. 14.
    Allen DG, Hutchins A-M, Hammet F et al (2002) Genetic aberrations detected by comparative genomic hybridisation in vulvar cancers. Br J Cancer 86(6):924–928PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Dehan E, Ben-Dor A, Liao W et al (2007) Chromosomal aberrations and gene expression profiles in non-small cell lung cancer. Lung Cancer 56(2):175–184PubMedCrossRefGoogle Scholar
  16. 16.
    Chang YC, Yeh K-T, Liu T-C, Chang J-G (2010) Molecular cytogenetic characterization of esophageal cancer detected by comparative genomic hybridization. J Clin Lab Anal 24(3):167–174PubMedCrossRefGoogle Scholar
  17. 17.
    Haverty PM, Hon LS, Kaminker JS, Chant J, Zhang Z (2009) High-resolution analysis of copy number alterations and associated expression changes in ovarian tumors. BMC Med Genom 2(1):21CrossRefGoogle Scholar
  18. 18.
    Sheu JJ-C, Lee C-H, Ko J-Y et al (2009) Chromosome 3p12. 3-p14. 2 and 3q26. 2-q26. 32 are genomic markers for prognosis of advanced nasopharyngeal carcinoma. Cancer Epidemiol Biomark Prev 18(10):2709–2716CrossRefGoogle Scholar
  19. 19.
    Linxweiler M, Linxweiler J, Barth M et al (2012) Sec62 bridges the gap from 3q amplification to molecular cell biology in non-small cell lung cancer. Am J Pathol 180(2):473–483PubMedCrossRefGoogle Scholar
  20. 20.
    Bochen F, Adisurya H, Wemmert S et al (2017) Effect of 3q oncogenes SEC62 and SOX2 on lymphatic metastasis and clinical outcome of head and neck squamous cell carcinomas. Oncotarget 8(3):4922PubMedCrossRefGoogle Scholar
  21. 21.
    Linxweiler M, Schick B, Zimmermann R (2017) Let’s talk about Secs: Sec61, Sec62 and Sec63 in signal transduction, oncology and personalized medicine. Signal Transduct Target Ther. 2:17002PubMedPubMedCentralCrossRefGoogle Scholar
  22. 22.
    Gerdes J, Lemke H, Baisch H, Wacker H-H, Schwab U, Stein H (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 133(4):1710–1715PubMedGoogle Scholar
  23. 23.
    Oberg TN, Kipp BR, Vrana JA et al (2010) Comparison of p16INK4a and ProEx C immunostaining on cervical ThinPrep cytology and biopsy specimens. Diagn Cytopathol 38(8):564–572.  https://doi.org/10.1002/dc.21251 PubMedCrossRefGoogle Scholar
  24. 24.
    Petry KU, Schmidt D, Scherbring S et al (2011) Triaging pap cytology negative, HPV positive cervical cancer screening results with p16/Ki-67 dual-stained cytology. Gynecol Oncol 121(3):505–509.  https://doi.org/10.1016/j.ygyno.2011.02.033 PubMedCrossRefGoogle Scholar
  25. 25.
    Schmidt D, Bergeron C, Denton KJ, Ridder R (2011) p16/ki-67 dual-stain cytology in the triage of ASCUS and LSIL Papanicolaou cytology. Cancer Cytopathol 119(3):158–166PubMedCrossRefGoogle Scholar
  26. 26.
    Van den Einden LCG, Grefte JMM, van der Avoort IAM, Vedder JEM, van Kempen LCLT, Massuger LFAG, de Hullu JA (2012) Cytology of the vulva: feasibility and preliminary results of a new brush. Br J Cancer 106(2):269–273.  https://doi.org/10.1038/bjc.2011.533 PubMedCrossRefGoogle Scholar
  27. 27.
    Jiménez-Ayala M, Jiménez-Ayala B (2002) Terminology for vulvar cytology based on the Bethesda system. Acta Cytol 46(4):645–650PubMedCrossRefGoogle Scholar
  28. 28.
    Bornstein J, Sideri M, Tatti S et al (2012) 2011 terminology of the vulva of the international federation for cervical pathology and colposcopy. J Low Genit Tract Dis 16(3):290–295PubMedCrossRefGoogle Scholar
  29. 29.
    Takacs FZ, Radosa JC, Bohle RM, Bochen F, Juhasz-Böss I, Solomayer EF, Schick B, Linxweiler M (2018) Sec62/Ki67 dual staining in cervical cytology specimens: a new marker for high-grade dysplasia. Arch Gynecol Obstet.  https://doi.org/10.1007/s00404-018-4981-4 PubMedCrossRefGoogle Scholar
  30. 30.
    Jiménez-Ayala M, Portillo BJ-A (2011) Glandular lesions of the vulva. Cytopathology of the glandular lesions of the female genital tract. In: Monographs in clinical cytology, vol 20. Karger Publishers, pp 77–86Google Scholar
  31. 31.
    Van der Avoort IAM, van der Laak JAWM, Paffen A et al (2007) MIB1 expression in basal cell layer: a diagnostic tool to identify premalignancies of the vulva. Mod Pathol 20(7):770–778PubMedCrossRefGoogle Scholar
  32. 32.
    Stewart C, Crook M (2014) Fascin and cyclin D1 immunoreactivity in non-neoplastic vulvar squamous epithelium, vulvar intraepithelial neoplasia and invasive squamous carcinoma: correlation with Ki67 and p16 protein expression. J Clin Pathol 67:319–325PubMedCrossRefGoogle Scholar
  33. 33.
    Bergeron C, Ikenberg H, Sideri M et al (2015) Prospective evaluation of p16/Ki-67 dual-stained cytology for managing women with abnormal Papanicolaou cytology: PALMS study results. Cancer Cytopathol 123(6):373–381PubMedCrossRefGoogle Scholar
  34. 34.
    Waldstrøm M, Christensen RK, Ornskov D (2013) Evaluation of p16INK4a/Ki-67 dual stain in comparison with an mRNA human papillomavirus test on liquid-based cytology samples with low-grade squamous intraepithelial lesion. Cancer Cytopathol 121(3):136–145PubMedCrossRefGoogle Scholar
  35. 35.
    Linxweiler M, Bochen F, Wemmert S et al (2015) Combination of p16INK4a/Ki67 immunocytology and HPV polymerase chain reaction for the noninvasive analysis of HPV involvement in head and neck cancer. Cancer Cytopathol 123(4):219–229PubMedCrossRefGoogle Scholar
  36. 36.
    Laara E, Day N, Hakama M (1987) Trends in mortality from cervical cancer in the Nordic countries: association with organised screening programmes. Lancet 329(8544):1247–1249CrossRefGoogle Scholar
  37. 37.
    Eva LJ (2012) Screening and follow up of vulval skin disorders. Best Pract Res Clin Obstet Gynaecol 26(2):175–188.  https://doi.org/10.1016/j.bpobgyn.2011.11.005 PubMedCrossRefGoogle Scholar
  38. 38.
    Dennerstein GJ (1968) The cytology of the vulva. J Obstet Gynaecol Br Commonw 75(6):603–609PubMedCrossRefGoogle Scholar
  39. 39.
    Nauth H, Schilke E (1982) Cytology of the exfoliative layer in normal and diseased vulvar skin: correlation with histology. Acta Cytol 26(3):269–283PubMedGoogle Scholar
  40. 40.
    Nauth H, Bӧger A (1982) New aspects of vulvar cytology. Acta Cytol 26(1):1–6PubMedGoogle Scholar
  41. 41.
    Bae-Jump VL, Bauer M, Van Le L (2007) Cytological evaluation correlates poorly with histological diagnosis of vulvar neoplasias. J Low Genit Tract Dis 11(1):8–11PubMedCrossRefGoogle Scholar
  42. 42.
    Levine T, Rolfe K, Crow J, Styles S, Perrett C, Maclean A, Reid W (2001) The use of cytospin monolayer technique in the cytological diagnosis of vulval and anal disease. Cytopathology 12(5):297–305PubMedCrossRefGoogle Scholar
  43. 43.
    Dennerstein G (2009) Vulvar cytology: useful or not? J Low Genit Tract Dis 13(3):192.  https://doi.org/10.1097/lgt.0b013e31818e159a (author reply 192) PubMedCrossRefGoogle Scholar
  44. 44.
    Rush D, Hyjek E, Baergen RN, Ellenson LH, Pirog EC (2005) Detection of microinvasion in vulvar and cervical intraepithelial neoplasia using double immunostaining for cytokeratin and basement membrane components. Arch Pathol Lab Med 129(6):747–753PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Ferenc Zoltan Takacs
    • 1
    Email author return OK on get
  • Julia Caroline Radosa
    • 1
  • Florian Bochen
    • 2
  • Ingolf Juhasz-Böss
    • 1
  • Erich-Franz Solomayer
    • 1
  • Rainer M. Bohle
    • 3
  • Georg-Peter Breitbach
    • 1
  • Bernard Schick
    • 2
  • Maximilian Linxweiler
    • 2
  1. 1.Department of Obstetrics and GynecologyUniversity of SaarlandHomburgGermany
  2. 2.Department of Otorhinolaryngology, Head and Neck SurgeryUniversity of SaarlandHomburgGermany
  3. 3.Department of General and Surgical PathologyUniversity of SaarlandHomburgGermany

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