Advertisement

Sec62/Ki67 dual staining in cervical cytology specimens: a new marker for high-grade dysplasia

  • Ferenc Zoltan TakacsEmail author
  • Julia Caroline Radosa
  • Rainer Maria Bohle
  • Florian Bochen
  • Ingolf Juhasz-Böss
  • Erich-Franz Solomayer
  • Bernard Schick
  • Maximilian Linxweiler
General Gynecology
  • 69 Downloads

Abstract

Purpose

In the previous studies, we demonstrated that Sec62 is essential for tumor cell migration, epithelial-to-mesenchymal transition, and intracellular stress tolerance. An increase in Sec62 expression correlated with an increase in cervical dysplasia severity in liquid-based cytology specimens. Ki67 is an established proliferation marker. Thus, in this study, we examined a method of Sec62/Ki67 dual staining for the detection of high-grade dysplasia and cancer in cervical liquid-based cytology specimens.

Methods

Sec62/Ki67 dual staining was performed on 100 cervical liquid-based cytology specimens. The staining results were correlated with cytological, immunocytological (p16/Ki67), colposcopic, and histological findings.

Results

All 56 (n = 56, 100%) cases of cervical intraepithelial neoplasia grade 3 and cervical cancer (CIN3+ lesions) were positive for Sec62/Ki67 staining, while low-grade lesions and normal cells were negative. Sec62/Ki67 staining was highly sensitive and specific for the detection of CIN2+ and CIN3+ lesions (94.37%; 100% and 100%; 84.09%, respectively).

Conclusions

Sec62/Ki67 dual-staining immunocytochemistry is a promising cytological tool for interpreting high-grade squamous lesions in cytological specimens and for assessing the risk of progression to cancer.

Keywords

Cervical cancer Cervical intraepithelial neoplasia Cytology Dual staining Immunocytochemistry Sec62 

Notes

Acknowledgements

The authors would like to gratefully acknowledge the excellent work, motivation, and enthusiasm of Mrs. Barbara Linxweiler and Mrs. Alice Kunz in the establishment of our dual-staining cytology method. We also want to thank our colposcopy team, namely, Mrs. Simona Baus, Mrs. Lisa Mees, Mrs. Frederike Schlaegel, Ms. Alexandra Sehr, and Mrs. Christina Linsler, for patient recruitment and for providing high-quality LBC specimens.

Author contributions

FZT: project development, data collection and analysis, manuscript writing. JCR: review and editing. FB: visualization, review and editing. RMB: review and editing. IJ-B: review and editing. EFS: review and editing. BS: review and editing. ML: project development, data analysis, review and editing.

Compliance with ethical standards

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or 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.

Conflict of interest

F. Z. Takacs and M. Linxweiler have a pending patent application for the Sec62/Ki67 immunocytochemical dual staining (application number: LU100824). All other authors declare no conflicts of interest.

References

  1. 1.
    Parkin DM, Stjernswärd J, Muir CS (1984) Estimates of the worldwide frequency of twelve major cancers. Bull World Health Organ 62:163–182PubMedPubMedCentralGoogle Scholar
  2. 2.
    Laara E, Day N, Hakama M (1987) Trends in mortality from cervical cancer in the Nordic countries: association with organised screening programmes. Lancet 329:1247–1249CrossRefGoogle Scholar
  3. 3.
    Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136:359–386CrossRefGoogle Scholar
  4. 4.
    Nanda K, McCrory DC, Myers ER, Bastian LA, Hasselblad V, Hickey JD, Matchar DB (2000) Accuracy of the papanicolaou test in screening for and follow-up of cervical cytologic abnormalities: a systematic review. Ann Intern Med 132:810–819CrossRefGoogle Scholar
  5. 5.
    Sano T, Oyama T, Kashiwabara K, Fukuda T, Nakajima T (1998) Immunohistochemical overexpression of p16 protein associated with intact retinoblastoma protein expression in cervical cancer and cervical intraepithelial neoplasia. Pathol Int 48:580–585CrossRefGoogle Scholar
  6. 6.
    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 119:158–166CrossRefGoogle Scholar
  7. 7.
    Petry KU, Schmidt D, Scherbring S, Luyten A, Reinecke-Lüthge A, Bergeron C, Kommoss F, Löning T, Ordi J, Regauer S, Ridder R (2011) Triaging Pap cytology negative, HPV positive cervical cancer screening results with p16/Ki-67 dual-stained cytology. Gynecol Oncol 121:505–509.  https://doi.org/10.1016/j.ygyno.2011.02.033 CrossRefPubMedGoogle Scholar
  8. 8.
    Bergeron C, Ikenberg H, Sideri M, Denton K, Bogers J, Schmidt D, Alameda F, Keller T, Rehm S, Ridder R (2015) Prospective evaluation of p16/Ki-67 dual-stained cytology for managing women with abnormal Papanicolaou cytology: PALMS study results. Cancer Cytopathol 123:373–381CrossRefGoogle Scholar
  9. 9.
    Shroyer KR, 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 Cytopathol 108:324–330CrossRefGoogle Scholar
  10. 10.
    Tosuner Z, Türkmen I, Arici S, Sӧnmez C, Turna S, Onaran Ö (2017) Immunocytoexpression profile of ProExC in smears interpreted as ASC-US, ASC-H, and cervical intraepithelial lesion. J Cytol 34:34–38CrossRefGoogle Scholar
  11. 11.
    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 121:136–145CrossRefGoogle Scholar
  12. 12.
    Heselmeyer K, Macville M, Schröck E, Blegen H, Hellström AC, Shah K, Auer G, Ried T (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 19:233–240CrossRefGoogle Scholar
  13. 13.
    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 210:412–419CrossRefGoogle Scholar
  14. 14.
    Linxweiler M, Bochen F, Schick B, Wemmert S, Al Kadah B, Greiner M, Hasenfus A, Bohle R-M, Juhasz-Bӧss I, Solomayer E-F, Takacs ZF (2016) Identification of SEC62 as a potential marker for 3q amplification and cellular migration in dysplastic cervical lesions. BMC Cancer 16:676CrossRefGoogle Scholar
  15. 15.
    Wright TC, Compagno J, Romano P, Grazioli V, Verma Y, Kershnar E, Tafas T, Kilpatrick MW (2015) Amplification of the 3q chromosomal region as a specific marker in cervical cancer. Am J Obstetr Gynecol 213(51):e1–e8Google Scholar
  16. 16.
    Allen DG, Hutchins A-M, Hammet F, White DJ, Scurry JP, Tabrizi SN, Garland SM, Armes JE (2002) Genetic aberrations detected by comparative genomic hybridisation in vulvar cancers. Br J Cancer 86:924–928CrossRefGoogle Scholar
  17. 17.
    Dehan E, Ben-Dor A, Liao W, Lipson D, Frimer H, Rienstein S, Simansky D, Krupsky M, Yaron P, Friedman E et al (2007) Chromosomal aberrations and gene expression profiles in non-small cell lung cancer. Lung Cancer 56:175–184CrossRefGoogle Scholar
  18. 18.
    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 24:167–174CrossRefGoogle Scholar
  19. 19.
    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 2:21CrossRefGoogle Scholar
  20. 20.
    Sheu JJ-C, Lee C-H, Ko J-Y, Tsao GS, Wu C-C, Fang C-Y, Tsai F-J, Hua C-H, Chen C-L, Chen J-Y (2009) Chromosome 3p12. 3–p14. 2 and 3q26. 2–q26. 32 are genomic markers for prognosis of advanced nasopharyngeal carcinoma. Cancer Epidemiol Prev Biomark 18:2709–2716CrossRefGoogle Scholar
  21. 21.
    Linxweiler M, Linxweiler J, Barth M, Benedix J, Jung V, Kim Y-J, Bohle RM, Zimmermann R, Greiner M (2012) Sec62 bridges the gap from 3q amplification to molecular cell biology in non-small cell lung cancer. Am J Pathol 180:473–483CrossRefGoogle Scholar
  22. 22.
    Bochen F, Adisurya H, Wemmert S, Lerner C, Greiner M, Zimmermann R, Hasenfus A, Wagner M, Smola S, Pfuhl T, Bozzato A, Kadah BA, Schick B, Linxweiler M (2017) Effect of 3q oncogenes SEC62 and SOX2 on lymphatic metastasis and clinical outcome of head and neck squamous cell carcinomas. Oncotarget 8:4922CrossRefGoogle Scholar
  23. 23.
    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 Targeted Ther 2:17002CrossRefGoogle Scholar
  24. 24.
    Hagerstrand D, Tong A, Schumacher SE, Ilic N, Shen RR, Cheung HW, Vazquez F, Shrestha Y, Kim SY, Giacomelli AO, Rosenbluh Joseph, 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 3:1044–1057CrossRefGoogle Scholar
  25. 25.
    Greiner M, Kreutzer B, Jung V, Grobholz R, Hasenfus A, Stöhr RF, Tornillo L, Dudek J, Stöckle M, Unteregger G, Kamradt J, Wullich B, Zimmermann R (2011) Silencing of the SEC62 gene inhibits migratory and invasive potential of various tumor cells. Int J Cancer 128:2284–2295.  https://doi.org/10.1002/ijc.25580 CrossRefPubMedGoogle Scholar
  26. 26.
    Linxweiler M, Schorr S, Schäuble N, Jung M, Linxweiler J, Langer F, Schäfers H-J, Cavalié A, Zimmermann R, Greiner M (2013) Targeting cell migration and the endoplasmic reticulum stress response with calmodulin antagonists: a clinically tested small molecule phenocopy of SEC62 gene silencing in human tumor cells. BMC Cancer 13:574CrossRefGoogle Scholar
  27. 27.
    Trunk MJ, Dallenbach-Hellweg G, Ridder R, Petry KU, Ikenberg H, Schneider V, von Knebel Doeberitz M (2004) Morphologic characteristics of p16INK4a-positive cells in cervical cytology samples. Acta Cytologica 48:771–782CrossRefGoogle Scholar
  28. 28.
    Lastra RR, Ou JJ, Reilly ME, Lawrence WD, Baloch ZW, Brooks JS, Barroeta JE (2014) Utility of ProExC and IMP3 immunocytochemical staining in atypical glandular cells of undetermined significance in liquid-based cervical cytology. Diagn Cytopathol 42:375–379CrossRefGoogle Scholar
  29. 29.
    Oberg TN, Kipp BR, Vrana JA, Bartholet MK, Fales CJ, Garcia R, McDonald AN, Rosas BL, Henry MR, Clayton AC (2010) Comparison of p16INK4a and ProEx C immunostaining on cervical ThinPrep cytology and biopsy specimens. Diagn Cytopathol 38:564–572.  https://doi.org/10.1002/dc.21251 CrossRefPubMedGoogle Scholar
  30. 30.
    Konishi I, Fujii S, Nonogaki H, Nanbu Y, Iwai T, Mori T (1991) Immunohistochemical analysis of estrogen receptors, progesterone receptors, Ki-67 antigen, and human papillomavirus DNA in normal and neoplastic epithelium of the uterine cervix. Cancer 68:1340–1350CrossRefGoogle Scholar
  31. 31.
    Dunton CJ, van Hoeven KH, Kovatich AJ, Oliver RE, Scacheri RQ, Cater JR, Carlson JA (1997) Ki-67 antigen staining as an adjunct to identifying cervical intraepithelial neoplasia. Gynecol Oncol 64:451–455CrossRefGoogle Scholar
  32. 32.
    McCredie MR, Sharples KJ, Paul C, Baranyai J, Medley G, Jones RW, Skegg DC (2008) Natural history of cervical neoplasia and risk of invasive cancer in women with cervical intraepithelial neoplasia 3: a retrospective cohort study. Lancet Oncol 9:425–434CrossRefGoogle Scholar
  33. 33.
    Habbema D, Weinmann S, Arbyn M, Kamineni A, Williams AE, de Kok MCM, I, van Kemenade F, Field TS, van Rosmalen J, Brown ML, (2017) Harms of cervical cancer screening in the United States and the Netherlands. Int J Cancer 140:1215–1222CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

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

Personalised recommendations