Mehmet Coşkun SALMAN, Koray ERGÜNAY, Alp USUBÜTÜN, Erdem KARABULUT, Ahmet PINAR, Sevgen Çelik ÖNDER, Aylin ALTAY, Gülendam BOZDAYI, Osman Selim BADUR, İpek TÜNEY, Kunter YÜCE

HPV types and E6/E7 mRNA expression in cervical samples from Turkish women with abnormal cytology in Ankara, Turkey

Background/aim: Human papillomaviruses have been established as a risk factor for invasive carcinoma of the uterine cervix. HPV E6/ E7 oncogene expression has recently emerged as a promising biomarker to determine the risk for progression to high-grade cervical lesions. The aim of this study was to evaluate HPV mRNA and DNA detection in samples with abnormal cytology. Materials and methods: Cervical specimens were obtained at the Department of Obstetrics and Gynecology via cervical brushes during January October 2011. Liquid-based cytology slides were evaluated according to the 2001 Bethesda System. Cytology specimens from a total of 81 women with abnormal cytology were included. Real-time PCR and NASBA assays were performed to detect HPV DNA and E6/E7 mRNA, respectively. Results: HPV DNA was identified in 73 samples (90.1%). HPV E6/E7 mRNA expression was observed in 45 samples (55.6%). A statistically significant difference was observed among cytological diagnosis groups. In 25 patients, a biopsy was performed during the follow-up. HPV DNA was detected in all of these patients. HPV E6/E7 expression was present only in CIN I III diagnosed patients. Conclusion: The E6/E7 mRNA test is a robust indicator of cytological atypia and correlates better with progressive lesions than DNA assays.

PDF

___

1. Zur Hausen H. Papillomaviruses causing cancer: evasion from host cell control in early events in carcinogenesis. J Natl Cancer Inst 2000; 92: 690-698.
2. de Villiers EM, Fauquet C, Broker TR, Bernard HU, Zur Hausen H. Classification of papillomaviruses. Virology 2004; 324: 17-27.
3. Schiffman MH, Bauer HM, Hoover RN, Glass AG, Cadell DM, Rush BB, Scott DR, Sherman ME, Kurman RJ, Wacholder S et al. Epidemiologic evidence showing that human papillomavirus infection causes most cervical intraepithelial neoplasia. J Natl Cancer Inst 1993; 85: 958-64.
4. Muñoz N, Bosch FX, de Sanjosé S, Herrero R, Castellsagué X, Shah KV, Snijders PJ, Meijer CJ; International Agency for Research on Cancer Multicenter Cervical Cancer Study Group. Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med 2003; 348: 518- 527.
5. Woodman CB, Collins S, Winter H, Bailey A, Ellis J, Prior P, Yates M, Rollason TP, Young LS. Natural history of cervical human papillomavirus infection in young women: a longitudinal cohort study. Lancet 2001; 357: 1831-1836.
6. Cuschieri KS, Whitley MJ, Cubie HA. Human papillomavirus type specific DNA and RNA persistence-implications for cervical disease progression and monitoring. J Med Virol 2004; 6: 1-6.
7. Schlecht NF, Kulaga S, Robitaille J, Ferreira S, Santos M, Miyamura RA, Duarte-Franco E, Rohan TE, Ferenczy A, Villa LL et al. Persistent human papillomavirus infection as a predictor of cervical intraepithelial neoplasia. JAMA 2001; 286: 3106-3114.
8. Nalliah S, Karikalan B, Kademane K. Multifaceted usage of HPV related tests and products in the management of cervical cancer a review. Asian Pac J Cancer Prev 2015; 16: 2145-2150.
9. Boulet GA, Horvath CA, Berghmans S, Bogers J. Human papillomavirus in cervical cancer screening: important role as biomarker. Cancer Epidemiol Biomarkers Prev 2008; 17: 810- 817.
10. Castle PE, Stoler MH, Wright TC, Sharma A, Wright TL, Behrens CM. Performance of carcinogenic human papillomavirus (HPV) testing and HPV16 or HPV18 genotyping for cervical cancer screening of women aged 25 years and older: a subanalysis of the ATHENA study. Lancet Oncol 2011; 12: 880-890.
11. Woodman CB, Collins S, Young LS. The natural history of cervical HPV infection: unresolved issues. Nat Rev Cancer 2007; 7: 11-22.
12. Zur Hausen H. Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer 2002; 2: 342-350.
13. Castle PE, Dockter J, Giachetti C, Garcia FAR, McCormick MK, Mitchell AL, Holladay EB, Kolk DP. A cross-sectional study of a prototype carcinogenic human papillomavirus E6/E7 messenger RNA assay for detection of cervical precancer and cancer. Clin Cancer Res 2007; 13: 2599-2605.
14. Coquillard G, Palao B, Patterson BK. Quantification of intracellular HPV E6/E7 mRNA expression increases the specificity and positive predictive value of cervical cancer screening compared to HPV DNA. Gynecol Oncol 2011; 120: 89-93.
15. Burger EA, Kornør H, Klemp M, Lauvrak V, Kristiansen IS. HPV mRNA tests for the detection of cervical intraepithelial neoplasia: a systematic review. Gynecol Oncol 2011; 120: 430-438.
16. Solomon D, Davey D, Kurman R, Moriarty A, OConnor D, Prey M, Raab S, Sherman M, Wilbur D, Wright T Jr et al. Forum Group Members, Bethesda 2001 Workshop: The 2001 Bethesda System: terminology for reporting results of cervical cytology. JAMA 2002; 287: 2114-2119.
17. Khunamornpong S, Settakorn J, Sukpan K, Srisomboon J, Suprasert P, Sriaunkgul S. Performance of HPV DNA testing with Hybrid Capture 2 in triaging women with minor cervical cytologic abnormalities (ASC-US/LSIL) in Northern Thailand. Asian Pac J Cancer Prev 2014; 15: 10961-10966.
18. Rossi PG, Benevolo M, Vocaturo A, Caraceni D, Ciccocioppo L, Frega A,Terranato I, Zappacosta R, French D, Rosini S. Prognostic value of HPV E6/E7 mRNA assay in women with negative colposcopy or CIN1 histology result: A follow-up study. PLoS ONE 2013; 8: e57600. doi:10.1371/journal.pone.0057600.
19. Discacciati MG, Silva ID, Villa LL, Reis L, Hayashi P, Costa MC, Rabelo-Santos SH, Zeferino LC. Prognostic value of DNA and mRNA E6/E7 of human papillomavirus in the evolution of cervical intraepithelial neoplasia grade 2. Biomarker Insights 2014; 9: 15-22.
20. Li N, Franceschi S, Howell-Jones R, Snijders PJF, Clifford GM. Human papillomavirus type distribution in 30,848 invasive cervical cancers worldwide: variation by geographical region, histological type and year of publication. Int J Cancer 2011; 128: 927-935.
21. Spathis A, Kottaridi C, Chranioti A, Meristoudis C, Chrelias C, Panayiotides IG, Paraskeviadis E, Karakitsos P. mRNA and DNA detection of human papillomaviruses in women of all ages attending two colposcopy clinics. PLoS ONE 2012; 7: e49205. doi:10.1371/journal.pone.0049205.
22. Tezcan S, Ozgur D, Ulger M, Aslan G, Gurses I, Serin MS, Giray BG, Dilek S, Emekdas G. HPV Genotype and E6/E7 oncogene expression in Turkish women with cervical cytology findings. Asian Pac J Cancer Prev 2014; 15: 3997-4003.
23. Broccolo F, Fusetti L, Rosini S, Caraceni D, Zappacosta R, Ciccocioppo L, Matteoli B, Halfon P, Malnati MS, CeccheriniNelli L. Comparison of oncogenic HPV type-specific viral DNA load and E6/E7 mRNA detection in cervical samples: results from a multicenter study. J Med Virol 2013; 85: 472-482.
24. Castro SP, Fernandez AI, Gonzales MJL, Diez MTS, Lama AC, Martin MJA, Mosquera MP, Lopez-Miragaya I, Estevez N, Pinon JT et al. Human papillomavirus (HPV) E6/E7 mRNA as a triage test after detection of HPV 16 and HPV 18 DNA. J Med Virol 2013; 85: 1063-1068.
25. Frega A, Lorenzon L, Giovagnoli MR, De Sangtis L, Fabriano V, Lukic A, Moscarini M, Torrisi MR, French D. Prognostic implication of high risk human papillomavirus E6 and E7 mRNA in patients with intraepithelial lesions of the cervix in relationship to age. Int J Immunopathol Pharmacol 2011; 24: 461-470.
26. Manavi M, Hudelist G, Fink-Retter A, Gschwantler-Kaulich D, Pischinger K, Czerwenka K. Human papillomavirus DNA integration and messenger RNA transcription in cervical lowand high-risk squamous intraepithelial lesions in Austrian women. Int J Gynecol Cancer 2008; 18: 285-294.
27. Persson, Elfström KM, Wendel SB, Weiderpass E, Andersson S. Triage of HR-HPV positive women with minor cytological abnormalities: a comparison of mRNA testing, HPV DNA testing, and repeat cytology using a 4-year follow-up of a populationbased study. PLoS ONE 2014; 9: e90023. doi:10.1371/ journal. pone.0090023.