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Open Access 01-12-2021 | Cervical Cancer | Research article

TOP2A/MCM2, p16^INK4a, and cyclin E1 expression in liquid-based cytology: a biomarkers panel for progression risk of cervical premalignant lesions

Authors: Oscar Del Moral-Hernández, Daniel Hernández-Sotelo, Luz del Carmen Alarcón-Romero, Miguel Angel Mendoza-Catalán, Eugenia Flores-Alfaro, Yaneth Castro-Coronel, Julio Ortiz-Ortiz, Marco Antonio Leyva-Vázquez, Carlos Ortuño-Pineda, Wendy Castro-Mora, Berenice Illades-Aguiar

Published in: BMC Cancer | Issue 1/2021

Abstract

Background

To improve the efficiency of early diagnosis systems for cervical cancer, the use of cellular and viral markers for identifying precancerous lesions with a greater probability to progress to cancer has been proposed. Several cellular proteins and markers of oxidative DNA damage have been suggested as possible biomarkers of cervical carcinogenesis; however, they have not been evaluated together. In this study, we analyzed the expression of the cellular markers p16^INK4a, Ki-67, CyclinE1, TOP2A/MCM2, and telomerase, as well as the DNA oxidative damage markers ROS and 8-OHdG. The analyses were performed in liquid-based cervical cytology samples or biopsies with premalignant lesions or cervical cancer diagnosis, with the purpose of selecting a panel of biomarkers that allow the identification of precursor lesions with greater risk of progression to cervical cancer.

Methods

We analyzed 1485 liquid-based cytology samples, including 239 non-squamous intraepithelial lesions (NSIL), 901 low-grade squamous intraepithelial lesions (LSIL), 54 high-grade squamous intraepithelial lesions (HSIL), and 291 cervical cancers (CC). The biomarkers were analyzed by immunocytochemistry and Human Papilloma Virus (HPV) genotyping with the INNO-LiPA genotyping Extra kit.

Results

We found that all tested cellular biomarkers were overexpressed in samples with high risk-HPV infection, and the expression levels increased with the severity of the lesion. TOP2A/MCM2 was the best biomarker for discriminating between LSIL and HSIL, followed by p16^INK4a and cyclinE1. Statistical analysis showed that TOP2A/MCM2 provided the largest explanation of HSIL and CC cases (93.8%), followed by p16^INK4a (91%), cyclin E1 (91%), Ki-67 (89.3%), and telomerase (88.9%).

Conclusions

We propose that the detection of TOP2A/MCM2, p16^INK4a and cyclin E1 expression levels is useful as a panel of biomarkers that allow identification of cervical lesions with a higher risk for progression to CC with high sensitivity and precision; this can be done inexpensively, in a single and non-invasive liquid-based cytology sample.

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Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015;136:E359–86.CrossRef

Comparetto C, Borruto F. Cervical cancer screening: A never-ending developing program. World J Clin Cases. 2015;3(7):614–24. https://doi.org/10.12998/wjcc.v3.i7.614.CrossRefPubMedPubMedCentral

Moody CA, Laimins LA. Human papillomavirus oncoproteins: pathways to transformation. Nat Rev Cancer. 2010;10:550–60.CrossRef

Crosbie EJ, Einstein MH, Franceschi S, Kitchener HC. Human papillomavirus and cervical cancer. Lancet. 2013;382(9895):889–99. https://doi.org/10.1016/S0140-6736(13)60022-7.CrossRefPubMed

Fahey MT, Irwig L, Macaskill P. Meta-analysis of pap test accuracy. Am J Epidemiol. 1995;141:680–9.CrossRef

Pinto AP, Degen M, Villa LL, Cibas ES. Immunomarkers in gynecologic cytology: the search for the ideal ‘biomolecular Papanicolaou test’. Acta Cytol. 2012;56:109–21.CrossRef

Capobianco G, Marras V, Wenger JM, Santeufemia DA, Ambrosini G, Lutzoni R, Dessole M, Cherchi PL. P16 immunostaining and HPV testing in histological specimens from the uterine cervix. Eur J Gynaecol Oncol. 2013;34(3):227–30.PubMed

Bahnassy AA, Zekri AR, Alam El-Din HM, Aboubakr AA, Kamel K, El-Sabah MT, et al. The role of cyclins and cyclins inhibitors in the multistep process of HPV-associated cervical carcinoma. J Egypt Natl Canc Inst. 2006;18:292–302.PubMed

Lv Q, Zhang J, Yi Y, Huang Y, Wang Y, Wang Y, et al. Proliferating cell nuclear antigen has an association with prognosis and risks factors of Cancer patients: a systematic review. Mol Neurobiol. 2016;53:6209–17.CrossRef

10.

Dixon EP, King LM, Nelson R, Simkins SG, Knapp SL, Brough GH, et al. Characterization and clinical validation of MCM2 and TOP2A monoclonal antibodies in the BD ProEx C assay: an immunoassay which detects aberrant S-phase induction in cervical tissue. J Immunol Methods. 2017;442:35–41.CrossRef

11.

Tosuner Z, Türkmen İ, Arıcı S, Sönmez C, Turna S, Onaran Ö. Immunocytoexpression profile of ProExC in smears interpreted as ASC-US, ASC-H, and cervical intraepithelial lesion. J Cytol. 2017;34:34–8. https://doi.org/10.4103/0970-9371.197605.CrossRefPubMedPubMedCentral

12.

Pańczyszyn A, Boniewska-Bernacka E, Głąb G. Telomeres and telomerase during human papillomavirus-induced carcinogenesis. Mol Diagn Ther. 2018;22(4):421–30. https://doi.org/10.1007/s40291-018-0336-x.CrossRefPubMedPubMedCentral

13.

Lewitowicz P, Nasierowska-Guttmejer A, Rokita W, Adamczyk-Gruszka O, Gluszek S, Chrapek M, et al. HPV genotyping and p16/Ki-67 test significantly improve detection rate of high-grade cervical squamous intraepithelial lesion. Arch Med Sci. 2019;16(1):87–93. https://doi.org/10.5114/aoms.2018.80697.CrossRefPubMedPubMedCentral

14.

Kaur G, Balasubramaniam SD, Lee YJ, Balakrishnan V, Oon CE. Minichromosome Maintenance Complex (MCM) Genes Profiling and MCM2 Protein Expression in Cervical Cancer Development. Asian Pac J Cancer Prev. 2019;20(10):3043–9. https://doi.org/10.31557/APJCP.2019.20.10.3043.CrossRefPubMedPubMedCentral

15.

Brown CA, Bogers J, Sahebali S, Depuydt CE, De Prins F, Malinowski DP. Role of protein biomarkers in the detection of high-grade disease in cervical cancer screening programs. J Oncol. 2012;2012:289315.PubMedPubMedCentral

16.

Moreno-Acosta P, Molano M, Morales N, Acosta J, González-Prieto C, Mayorga D, et al. hTERT protein expression in cytoplasm and nucleus and its association with HPV infection in patients with cervical cancer. Cancer Genomics Proteomics. 2020;17(5):615–25. https://doi.org/10.21873/cgp.20218.CrossRefPubMedPubMedCentral

17.

Jelić M, Mandić A, Kladar N, Sudji J, Božin B, Srdjenović B, et al. Lipid peroxidation, Antioxidative defense and level of 8-hydroxy-2-deoxyguanosine in cervical Cancer patients. J Med Biochem. 2018;37(3):336–45. https://doi.org/10.1515/jomb-2017-0053.CrossRefPubMedPubMedCentral

18.

Illades-Aguiar B, Alarcon-Romero Ldel C, Antonio-Vejar V, Zamudio-Lopez N, SalesLinares N, Flores-Alfaro E, et al. Prevalence and distribution of human papillomavirus types in cervical cancer, squamous intraepithelial lesions, and with no intraepithelial lesions in women from Southern Mexico. Gynecol Oncol. 2010;117:291–6.CrossRef

19.

Tokunaga H, Shimada M, Ishikawa M, Yaegashi N. TNM classification of gynaecological malignant tumours, eighth edition: changes between the seventh and eighth editions. Jpn J Clin Oncol. 2019;49(4):311–20. https://doi.org/10.1093/jjco/hyy206.CrossRefPubMed

20.

Davis LG, Kuehl WM, Battey JF. Basic methods in molecular biology. 2nd ed. Norwalk: Appleton & Lange; 1994.

21.

Kleter B, van Doorn LJ, Schrauwen L, Molijn A, Sastrowijoto S, ter Schegget J, et al. Development and clinical evaluation of a highly sensitive PCR-reverse hybridization line probe assay for detection and identification of anogenital human papillomavirus. J Clin Microbiol. 1999;37:2508–17.CrossRef

22.

Tagle DK, Sotelo DH, Illades-Aguiar B, Leyva-Vazquez MA, Alfaro EF, Coronel YC, et al. Expression of E6, p53 and p21 proteins and physical state of HPV16 in cervical cytologies with and without low grade lesions. Int J Clin Exp Med. 2014;7:186–93.PubMedPubMedCentral

23.

Zubillaga-Guerrero MI, Illades-Aguiar B, Leyva-Vazquez MA, Flores-Alfaro E, Castaneda-Saucedo E, Munoz-Valle JF, et al. The integration of HR-HPV increases the expression of cyclins A and E in cytologies with and without low-grade lesions. J Cytol. 2013;30:1–7.CrossRef

24.

Gargano JW, Nisenbaum R, Lee DR, Ruffin MT, Steinau M, Horowitz IR, et al. Age group differences in human papillomavirus types and cofactors for cervical intraepithelial neoplasia 3 among women referred to colposcopy. Cancer Epidemiol Biomarkers Prev. 2012;21:111–21.CrossRef

25.

Illades-Aguiar B, Cortes-Malagon EM, Antonio-Vejar V, Zamudio-Lopez N. Alarcon Romero Ldel C, Fernandez-Tilapa G, et al. cervical carcinoma in southern Mexico: human papillomavirus and cofactors. Cancer Detect Prev. 2009;32:300–7.CrossRef

26.

Guardado-Estrada M, Juarez-Torres E, Roman-Bassaure E, Medina-Martinez I, Alfaro A, Benuto RE, et al. The distribution of high-risk human papillomaviruses is different in young and old patients with cervical cancer. PLoS One. 2014;9:e109406.CrossRef

27.

de Sanjose S, Quint WG, Alemany L, Geraets DT, Klaustermeier JE, Lloveras B, et al. Human papillomavirus genotype attribution in invasive cervical cancer: a retrospective cross-sectional worldwide study. Lancet Oncol. 2010;11:1048–56.CrossRef

28.

Guan P, Howell-Jones R, Li N, Bruni L, de Sanjose S, Franceschi S, et al. Human papillomavirus types in 115,789 HPV-positive women: a meta-analysis from cervical infection to cancer. Int J Cancer. 2012;131:2349–59.CrossRef

29.

Bruni L, Barrionuevo-Rosas L, Serrano B, Brotons M, Cosano R, Muñoz J. ICO information Centre on HPV and Cancer (HPV information Centre). Human Papillomavirus and related diseases in Mexico. Summary Report 2014. https://www.hpvcentre.net/statistics/reports/MEX.pdf. Accessed 4 Nov 2020.

30.

Chen EY, Tran A, Raho CJ, Birch CM, Crum CP, Hirsch MS. Histological ‘progression’ from low (LSIL) to high (HSIL) squamous intraepithelial lesion is an uncommon event and an indication for quality assurance review. Mod Pathol. 2010;23:1045–51.CrossRef

31.

Yang QC, Zhu Y, Liou HB, Zhang XJ, Shen Y, Ji XH. A cocktail of MCM2 and TOP2A, p16INK4a and Ki-67 as biomarkers for the improved diagnosis of cervical intraepithelial lesion. Pol J Pathol. 2013;64:21–7.CrossRef

32.

Branca M, Ciotti M, Santini D, Di Bonito L, Giorgi C, Benedetto A, et al. p16 (INK4a) expression is related to grade of CIN and high-risk human papillomavirus but does not predict virus clearance after conization or disease outcome. Int J Gynecol Pathol. 2004;23:354–65.CrossRef

33.

Han Q, Guo H, Geng L, Wang Y. p16/Ki-67 dual-stained cytology used for triage in cervical cancer opportunistic screening. Chin J Cancer Res. 2020;32(2):208–17. https://doi.org/10.21147/j.issn.1000-9604.2020.02.08.9.CrossRefPubMedPubMedCentral

34.

Liao GD, Sellors JW, Sun HK, Zhang X, Bao YP, Jeronimo J, et al. p16INK4A immunohistochemical staining and predictive value for progression of cervical intraepithelial neoplasia grade 1: a prospective study in China. Int J Cancer. 2014;134(7):1715–24. https://doi.org/10.1002/ijc.28485 Epub 2013 Oct 12.CrossRefPubMed

35.

Galgano MT, Castle PE, Atkins KA, Brix WK, Nassau SR, Stoler MH. Using biomarkers as objective standards in the diagnosis of cervical biopsies. Am J Surg Pathol. 2010;34:1077–87.CrossRef

36.

Zejnullahu VA, A Zejnullahu VA, Josifovska S, Vukovik N, Pakovski K, Panov S. Correlation of hTERT expression with cervical cytological abnormalities and human papillomavirus infection. Pril (Makedon Akad Nauk Umet Odd Med Nauki). 2017;38(3):143–51. https://doi.org/10.2478/prilozi-2018-0015.CrossRef

37.

Romano G, Sgambato A, Mancini R, Capelli G, Giovagnoli MR, Flamini G, et al. 8-hydroxy-2′-deoxyguanosine in cervical cells: correlation with grade of dysplasia and human papillomavirus infection. Carcinogenesis. 2000;21(6):1143–7.CrossRef

38.

Visalli G, Riso R, Facciolà A, Mondello P, Caruso C, Picerno I, et al. Higher levels of oxidative DNA damage in cervical cells are correlated with the grade of dysplasia and HPV infection. J Med Virol. 2016;88(2):336–44. https://doi.org/10.1002/jmv.24327.CrossRefPubMed

39.

Orang'o EO, Were E, Rode O, Muthoka K, Byczkowski M, Sartor H, et al. Novel concepts in cervical cancer screening: a comparison of VIA, HPV DNA test and p16 INK4a/Ki-67 dual stain cytology in Western Kenya. Infect Agent Cancer. 2020;15:57. https://doi.org/10.1186/s13027-020-00323-6.CrossRefPubMedPubMedCentral

40.

Hu Y, Hong Z, Gu L, Xie L, Yang B, Dai H, et al. Evaluation of p16/Ki-67 dual-stained cytology in triaging HPV-positive women during cervical Cancer screening. Cancer Epidemiol Biomark Prev. 2020;29(6):1246–52. https://doi.org/10.1158/1055-9965.EPI-19-1180.CrossRef

Title: TOP2A/MCM2, p16INK4a, and cyclin E1 expression in liquid-based cytology: a biomarkers panel for progression risk of cervical premalignant lesions
Authors: Oscar Del Moral-Hernández
Daniel Hernández-Sotelo
Luz del Carmen Alarcón-Romero
Miguel Angel Mendoza-Catalán
Eugenia Flores-Alfaro
Yaneth Castro-Coronel
Julio Ortiz-Ortiz
Marco Antonio Leyva-Vázquez
Carlos Ortuño-Pineda
Wendy Castro-Mora
Berenice Illades-Aguiar
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Cervical Cancer
Cervical Cancer
Biomarkers
Published in: BMC Cancer / Issue 1/2021
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-020-07740-1

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