The role of p16 / Ki-67 dual staining in HPV positive and negative women in the early diagnosis of cervical precancerous lesions: Cytology, colposcopy and conization protocol

Aim: The aim of this study was to evaluate the relationship between p16 / Ki-67 dual staining used for the definition of precancerous cervical lesions with histological results and HPV positivity. Materials and Method: This study is a cross-sectional study of 468 patients who were followed up in our center with the diagnosis of cervical intraepithelial neoplasia between 2016 and 2019 using the cytology, colposcopic biopsy and conization results, HPV test and p16 / Ki-67 dual staining results. SPSS 22 program was used in the analysis of the data. In the analysis of qualitative data, chi-square test and binary logistic regression analysis were used. The compatibility of both models with bilateral logistic regression test was good (omnibus test < 0.001). The correct estimate percentage of the model is 71.4% and 80.3%. p <0.05 is considered important. Results: In the binary logistic regression test established between HPV types and p16 / Ki-67 dual staining positivity, other high risk HPV types, HPV 16, 18 and 16-18, increased p16 / Ki-67 positivity ratio in this order. In the binary logistic regression test established between abnormal cytology and p16 / Ki-67 dual staining positivity in the colposcopy results, there was p16 / Ki-67 positivity increasing in proportion with the degree of HGSIL lesions. Conclusion: This study created with cytology / colposcopy and LEEP conization protocol shows that; in effective screening for early diagnosis and treatment in cervical cancer, p16 / Ki-67 biomarkers can be used effectively.

p16/Ki-67 dual staining; HGSIL; cytology; colposcopy; LEEP conization; HPV posivite

[1] Arbyn M., Weiderpass E., Bruni L., de Sanjosé S., Saraiya M., Ferlay J., et al.:. ”Estimates of incidence and mortality of cervical cancer in 2018: a worldwide analysis”. Lancet Glob Health. 2020, 8, e191.

[2] Saslow D., Solomon D., Lawson H.W., Killackey M., Kulasingam S.L., Cain J.M., et al.:. ”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”. Journal of lower genital tract disease. 2012, 16, 175.

[3] Castle P.E., Fetterman B., Thomas Cox J., Shaber R., Poitras N., Lorey T., et al.:. ”The Age-Specific Relationships of Abnormal Cytology and Human Papillomavirus DNA Results to the Risk of Cervical Precancer and Cancer”. Obstetrics & Gynecology. 2010, 116, 76.

[4] de Sanjose S., Quint W.G.V., Alemany L., Geraets D.T., Klauster- meier J.E., Lloveras B., et al.:. ”Human papillomavirus genotype attribution in invasive cervical cancer: a retrospective cross-sectional worldwide study”. The Lancet Oncology. 2010, 11, 1048.

[5] Kjær S.K., Frederiksen K., Munk C.., Iftner T.: ”Long-term absolute risk of cervical intraepithelial neoplasia grade 3 or worse following human papillomavirus infection: role of persistence”. J Natl Cancer Inst. 2010, 102, 1478.

[6] Liao G.-D., Sellors J.W., Sun H.-K., Zhang X., Bao Y.-P., Jeronimo J., et al.:. ”p16INK4Aimmunohistochemical staining and predictive value for progression of cervical intraepithelial neoplasia grade 1: A prospective study in China”. International Journal of Cancer. 2013, 134, 1715.

[7] Gupta N., Srinivasan R.., Rajwanshi A.: ”Functional biomarkers in cervical precancer: An overview”. Diagnostic Cytopathology. 2009, NA.

[8] el-All H.S.A., Refaat A.., Dandash K.: ”Prevalence of cervical neoplastic lesions and Human Papilloma Virus infection in Egypt: National Cervical Cancer Screening Project”. Infect Agent Cancer. 2007, 2, 12.

[9] Ikenberg H., Bergeron C., Schmidt D., Griesser H., Alameda F., Angeloni C., et al.:. ”Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study”. J Natl Cancer Inst. 2013, 105, 1550.

[10] Petry K.U., Schmidt D., Scherbring S., Luyten A., Reinecke-Lüthge A., Bergeron C., et al.:. ”Triaging Pap cytology negative, HPV positive cervical cancer screening results with p16/Ki-67 Dual-stained cytology”. Gynecologic Oncology. 2011, 121, 505.

[11] Wentzensen N., Schwartz L., Zuna R.E., Smith K., Mathews C., Gold M.A., et al.:. ”Performance of p16/Ki-67 immunostaining to detect cervical cancer precursors in a colposcopy referral population”. Clin Cancer Res. 2012, 18, 4154.

[12] Lesnikova I., Lidang M., Hamilton-Dutoit S.., Koch J.: ”p16 as a diagnostic marker of cervical neoplasia: a tissue microarray study of 796 archival specimens”. Diagn Pathol. 2009, 4, 22.

[13] Ancuta E., Ancuta C., Cozma L.G., Iordache C., Anghelache-Lupascu I., Anton E., et al.:. ”Tumor biomarkers in cervical cancer: focus on Ki-67 proliferation factor and E-cadherin expression”. 2009, 50, 413.

[14] Sadri Nahand J., Moghoofei M., Salmaninejad A., Bahmanpour Z., Karimzadeh M., Nasiri M., et al.:. ”Pathogenic role of exosomes and microRNAs in HPV-mediated inflammation and cervical cancer: A review”. International journal of cancer. 2020, 146, 305.

[15] Uijterwaal M.H., Polman N.J., Witte B.I., van Kemenade F.J., Ri- jkaart D., Berkhof J., et al.:. ”Triaging HPV-positive women with normal cytology by p16/Ki-67 dual-stained cytology testing: Baseline and longitudinal data”. International Journal of Cancer. 2014, 136, 2361.

[16] Uijterwaal M.H., Witte B.I., Van Kemenade F.J., Rijkaart D., Ridder R., Berkhof J., et al.:. ”Triaging borderline/mild dyskaryotic Pap cytology with p16/Ki-67 dual-stained cytology testing: cross-sectional and longitudinal outcome study”. British journal of cancer. 2014, 110, 1579.

[17] Nahand J.S., Taghizadeh-boroujeni S., Karimzadeh M., Borran S., Pourhanifeh M.H., Moghoofei M., et al.:. ”microRNAs: New prognostic, diagnostic, and therapeutic biomarkers in cervical cancer”. Journal of Cellular Physiology. 2019, 234, 17064.

[18] Ghasemi F., Shafiee M., Banikazemi Z., Pourhanifeh M.H., Khan- babaei H., Shamshirian A., et al.:. ”Curcumin inhibits NF-kB and Wnt/β-catenin pathways in cervical cancer cells”. Pathology - Research and Practice. 2019, 215, 152556.

[19] Shafabakhsh R., Reiter R.J., Mirzaei H., Teymoordash S.N.., Asemi Z.: ”Melatonin: A new inhibitor agent for cervical cancer treatment”. Journal of Cellular Physiology. 2019, 234, 21670.

[20] Wentzensen N., Fetterman B., Castle P.E., Schiffman M., Wood S.N., Stiemerling E., et al.:. ”p16/Ki-67 Dual Stain Cytology for Detection of Cervical Precancer in HPV-Positive Women”. J Natl Cancer Inst. 2015, 107, djv257.

[21] Zhong P., Li J., Gu Y., Liu Y., Wang A., Sun Y., et al.:. ”P16 and Ki- 67 expression improves the diagnostic accuracy of cervical lesions but not predict persistent high risk human papillomavirus infection with CIN1”. 2015, 8, 2979.

[22] Schmidt D., Bergeron C., Denton K.J.., Ridder R.: ”p16/ki-67 dualstain cytology in the triage of ASCUS and LSIL Papanicolaou cytology”. Cancer Cytopathology. 2011, 119, 158.

[23] Leite P., Tafuri L., Costa M., Lima M.., Simões R.: ”Evaluation of the p16 and Ki-67 Biomarkers as Predictors of the Recurrence of Premalignant Cervical Cancer Lesions after LEEP Conization”. Revista Brasileira de Ginecologia e Obstetrícia / RBGO Gynecology and Obstetrics. 2017, 39, 288.

[24] Yu L.-L., Chen W., Lei X.-Q., Qin Y., Wu Z.-N., Pan Q.-J., et al.:. ”Evaluation of p16/Ki-67 dual staining in detection of cervical precancer and cancers: a multicenter study in China”. Oncotarget. 2016, 7, 21181.