Abstract
Several biomarkers are in use to improve the sensitivity and specificity of cervical cancer screening. Previously, increased expression of tight junction protein claudin-1 (CLDN1) was detected in premalignant and malignant cervical lesions and applied for cytology screening. To improve the specificity, a double immunoreaction with CLDN1/Ki67 was developed in the recent study. Parallel p16/Ki67 (CINtec® PLUS) and CLDN1/Ki67 dual-stained cytology and histology were performed and compared. p16/Ki67 immunoreaction showed positivity in 317 out of 1596 smears with negativity in 1072 and unacceptable reactions in 207 samples. CLDN1/Ki67 dual staining was positive in 200 of 1358 samples, negative in 962, whereas 196 smears could not be evaluated due to technical reasons. Considering the high-grade squamous intraepithelial lesion cytology as gold standard, sensitivity of CLDN1/Ki67 reaction was 76%, specificity was 85.67%, while for p16/Ki67 sensitivity was 74% and specificity was 81.38%. Comparison of CLDN1/Ki67 and p16/Ki67 dual stainings showed the results of the two tests not to be significantly different. Analysing histological slides from 63 cases, the results of the two tests agreed perfectly. As conclusion the sensitivity and specificity proved to be similar using p16/Ki67 and CLDN1/Ki67 double immunoreactions both on LBC samples and on histological slides.
Similar content being viewed by others
References
Hillemans P, Soergel P, Hertel H, Jentschke M (2016) Epidemiology and early detection of cervical cancer. Oncol Res Treat 39:1–6
Sawaya GF, Smith-McCune K (2016) Cervical cancer screening. Obstet Gynecol 127:459–467
Lees BF, Erickson BK, Huh WK (2016) Cervical cancer screening: evidence behind the guidelines. Am J Obstet Gynecol 214:438–443
Ocque R, Austin RM (2016) Follow-up of women with negative pap test results and abnormal clinical signs or symptoms. Am J Clin Pathol 145:560–567
Ikenberg H, Bergeron C, Schmidt D, Griesser H, Alameda F, Angeloni C, Bogers J, Dachez R, Denton K, Hariri J, Keller T, von Knebel Doeberitz M, Neumann HH, Puig-Tintore LM, Sideri M, Rehm S, Ridder R, PALMS Study Group (2013) Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study. J Natl Cancer Inst 105:1550–1557
Wentzensen N, Fetterman B, Castle PE, Schiffman M, Wood SN, Stiemerling E, Tokugawa D, Bodelon C, Roitras N, Lorey T, Kinney W (2015) p16/Ki-67 dual stain cytology for detection of cervical precancer in HPV-positive women. J Natl Cancer Inst 107:1–8
Cuzick J, Clavel C, Petry KU, Meijer CJ, Hoyer H, Ratnam S, Szarewski A, Birembaut P, Kulasingam S, Sasieni P, Iftner T (2006) Overview of the European and north American studies on HPV testing in primary cervical cancer screening. Int J Cancer 119:1095–1101
Plummer M, Schiffman M, Castle PE, Maucort-Boulch D, Wheeler CM, ALTS Group (2007) A 2-year prospective study of human papillomavirus persistence among women with a cytological diagnosis of atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesion. J Infect Dis 195:1582–1589
Benczik M, Galamb Á, Koiss R, Kovács A, Járay B, Székely T, Szekerczés T, Schaff Z, Sobel G, Jeney C (2016) Claudin-1 as a biomarker of cervical cytology and histology. Pathol Oncol Res 22:179–188
Bergeron C, Ikenberg H, Sideri M, Denton K, Bogers J, Schmidt D, Alameda F, Keller T, Rehm S, Ridder R, PALMS Study Group (2015) Prospective evaluation of p16/Ki-67 dual-stained cytology for managing women with abnormal Papanicolaou cytology: PALMS study results. Cancer Cytopathol 123:373–381
Kocsis A, Takács T, Jeney C, Schaff Z, Koiss R, Járay B, Sobel G, Pap K, Székely I, Ferenci T, Lai HC, Nyíri M, Benczik M (2017) Performance of a new HPV and biomarker assay in management of hrHPV positive women: Subanalysis of the ongoing multicenter TRACE Clinical Trial (n>6,000) to evaluate POU4F3 as a potential biomarker of cervical precancer and cancer. Int J Cancer 140:1119–1133
Sobel G, Páska C, Szabó I, Kiss A, Kádár A, Schaff Z (2005) Increased expression of claudins in cervical squamous intraepithelial neoplasia and invasive carcinoma. Hum Pathol 36:162–169
Szabo I, Kiss A, Schaff Z, Sobel G (2009) Claudins as diagnostic and prognostic markers in gynecological cancer. Histol Histopathol 24:1607–1615
Lee JW, Lee SJ, Seo J, Song SY, Ahn G, Park CS, Lee JH, Kim BG, Bae DS (2005) Increased expressions of claudin-1 and claudin-7 during the progression of cervical neoplasia. Gynecol Oncol 97:53–59
Lal-Nag M, Morin PJ (2009) The claudins. Genome Biol 10:235
Nayar R, Wilbur DC (2015) The pap test and Bethesda 2014. Cancer Cytopathol 123:271–281
Solomon D (2015) Foreword. In: Nayar F, Wilbur DC (ed) The Bethesda system for reporting cervical cytology. Definitions, criteria, and explanatory notes, 3rd ed. Springer International Publishing Switzerland, Cham, pp v-vii
Wentzensen N, von Knebel Doeberitz M (2007) Biomarkers in cervical cancer screening. Dis Markers 23:315–330
Lee S, Rose MS, Sahasrabuddhe VV, Zhao R, Duggan MA (2017) Tissue-based immunohistochemical biomarker accuracy in the diagnosis of malignant glandular lesions of the uterine cervix: a systematic review of the literature and meta-analysis. Int J Gynecol Pathol 36:310–322
Polman NJ, Uijterwaal MH, Witte BI, Berkhof J, van Kemenade FJ, Spruijt JW, van Baal WM, Graziosi PG, van Dijken DK, Verheijen RH, Helmerhorst TJ, Steenbergen RD, Heideman DA, Ridder R, Snijders PJ, Meijer CJ (2017) Good performance of p16/ki-67 dual-stained cytology for surveillance of women treated for high-grade CIN. Int J Cancer 140:423–430
Yu LL, Guo HQ, Lei XQ, Qin Y, Wu ZN, Kang LN, Zhang X, Qiao YL, Chen W (2016) p16/Ki-67 co-expression associates high risk human papillomavirus persistence and cervical histopathology: a 3-year cohort study in China. Oncotarget 7:64810–64819
Possati-Resende JC, Fregnani JHTG, Kerr LM, Mauad EC, Longatto-Filho A, Scapulatempo-Neto C (2015) The accuracy of p16/Ki67 and HPV test in the detection of CIN2/3 in women diagnosed with ASC-US or LSIL. PLoS One 10:e0134445
Furuse M, Fujita K, Hiiragi T, Fujimoto K, Tsukita S (1998) Claudin-1 and -2: novel integral membrane proteins localizing at tight junctions with no sequence similarity to occludin. J Cell Biol 141:1539–1550
Morin PJ (2005) Claudin proteins in human cancer: promising new targets for diagnosis and therapy. Cancer Res 65:9603–9606
Soini Y (2005) Expression of claudins 1, 2, 3, 4, 5 and 7 in various types of tumours. Histopathology 46:551–560
Cunniffe C, Brankin B, Lambkin H, Ryan F (2014) The role of claudin-1 and claudin-7 in cervical tumorigenesis. Anticancer Res 34:2851–2857
Akimoto T, Takasawa A, Murata M, Kojima Y, Takasawa K, Nojima M, Aoyama T, Hiratsuka Y, Ono Y, Tanaka S, Osanai M, Hasegawa T, Saito T, Sawada N (2016) Analysis of the expression and localization of tight junction transmembrane proteins, claudin-1, −4, −7, occludin and JAM-A, in human cervical adenocarcinoma. Histol Histopathol 31:921–931
Győrffy H, Holczbauer Á, Nagy P, Szabó Z, Kupcsulik P, Páska C, Papp J, Schaff Z, Kiss A (2005) Claudin expression in Barrett’s esophagus and adenocarcinoma. Virchows Arch 447:961–968
Tőkés AM, Kulka J, Paku S, Szik Á, Páska C, Novák PK, Szilák L, Kiss A, Bögi K, Schaff Z (2005) Claudin-1, −3 and −4 proteins and mRNA expression in benign and malignant breast lesions: a research study. Breast Cancer Res 7:R296–R305
Lódi C, Szabó E, Holczbauer Á, Batmunkh E, Szíjártó A, Kupcsulik P, Kovalszky I, Paku S, Illyés G, Kiss A, Schaff Z (2006) Claudin-4 differentiates biliary tract cancers from hepatocellular carcinomas. Mod Pathol 19:460–469
Szász AM, Nyírády P, Majoros A, Szendrői A, Szűcs M, Székely E, Tőkés AM, Romics I, Kulka J (2010) Beta-catenin expression and claudin expression pattern as prognostic factors of prostatic cancer progression. BJU Int 105:716–722
English DP, Santin AD (2013) Claudins overexpression in ovarian cancer: potential targets for Clostridium Perfringens enterotoxin (CPE) based diagnosis and therapy. Int J Mol Sci 14:10412–10437
Vázquez-Ortíz G, Ciudad CJ, Pina P, Vazquez K, Hidalgo A, Latorre B, Garcia JA, Salamanca F, Peralta-Rodriguez R, Rangel A, Salcedo M (2005) Gene identification by cDNA arrays in HPV-positive cervical cancer. Arch Med Res 36:448–458
Funding
This work was supported by grant #NKFP_07_2-SPE-SAFE, Jedlik Ányos 2. subprogram KMR_12–1–2012-0032 by the Hungarian National Research and Development Fund and grant #OTKA PD105019 by the Hungarian National Research Foundation.
Author information
Authors and Affiliations
Contributions
TSz performed and evaluated the immunreactions and statistics, ÁG and AM collected the clinical samples and data, BJ and AKi evaluated the cytology, ZsS and GS designed and coordinated the study and wrote the manuscript. AKo and MB are clinical experts of the TRACE clinical study (Cellcall). TT is the inventor of the HPV detection technology applied in the CONFIDENCE HPV™ (NEUMANN Diagnostics) testing. CsJ was former head of the TRACE clinical study (Cellcall). ZsS accepts full responsibility for the work. All authors have read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare that they have no conflict of interest with respect to the research, authorship, and/or publication of this article.
AKo is an employee of Cellcall and NEUMANN. MB receives consultancy fees from Cellcall and NEUMANN, she is an employee of SYNLAB GenoID Laboratory, which participated in the TRACE clinical study (all related expenses were charged to Cellcall and NEUMANN). TT is a part time employee of Cellcall, he receives consultancy fees from NEUMANN, he is minority owner of NEUMANN, the inventor of the HPV detection technology described in the manuscript covering which Cellcall submitted a patent application (license now owned by NEUMANN Diagnostics Ltd.); he is a part time employee of SYNLAB GenoID Laboratory. CsJ was a former employee of Cellcall. MNy is an employee of Cellcall and NEUMANN.
Rights and permissions
About this article
Cite this article
Szekerczés, T., Galamb, Á., Kocsis, A. et al. Dual-Stained Cervical Cytology and Histology with Claudin-1 and Ki67. Pathol. Oncol. Res. 25, 477–486 (2019). https://doi.org/10.1007/s12253-018-0384-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12253-018-0384-x