Preferences help
enabled [disable] Abstract
Number of results
2017 | 64 | 1 | 143-149
Article title

HPV16 E6 polymorphism and physical state of viral genome in relation to the risk of cervical cancer in women from the south of Poland

Title variants
Languages of publication
The aim of this study was to analyse the correlation between HPV16 E6 variants and the physical status of viral genome (integrated, mixed, episomal) among patients with cervical cancer (n=40) and low-grade squamous intraepithelial lesions - LSIL (n=40). The study was performed on 80 HPV16 positive samples. HPV16 E6 variants were identified using PCR and DNA sequencing. Nucleotide sequences of E6 were compared with the prototype sequence (EUR-350T). The physical state of HPV DNA was determined as the ratio of E2/E6 copy number per cell. Twelve different intratypic variants were identified as belonging to European (in 77 samples) and North-American 1 (in 3 samples) sublineages. The most prevalent non-synonymous variant was EUR-350G, which occurred with similar frequency in cervical cancer and LSIL. The frequencies of additional mutations in variants with EUR-350T or EUR-350G sequences differed significantly. For the first time, missense mutations G122A, C153T and G188A were discovered in EUR-350G variant. The integrated viral genome was predominant in women with cervical cancer. The EUR-350T prototype and EUR-350G without additional mutations variants were prevalent in cervical cancer samples with the HPV16 characterized by integrated DNA. In summary, European variants of HPV16 E6 dominated in both cancer and LSIL group. The presence of EUR-350G favoured the occurrence of additional nucleotide changes. We showed that nucleotide changes occur significantly more often in the mixed form of viral DNA and in LSIL group and that the variants without additional mutations may promote integration of HPV16 genome.
Physical description
  • Department of Virology, Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Department of Virology, Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Department of Gynecological Oncology, Maria Skłodowska-Curie Memorial Cancer Centre and Institute of Oncology, Cracow Branch, Kraków, Poland
  • Department of Virology, Chair of Microbiology, Jagiellonian University Medical College, Kraków, Poland
  • Andersson S, Alemi M, Rylander E, Strand A, Larsson B, Sällström J, Wilander E (2000) Uneven distribution of HPV 16 E6 prototype and variant (L83V) oncoprotein in cervical neoplastic lesions. Br J Cancer 83: 307-310. doi: 10.1054/bjoc.2000.1247.
  • Bernard HU, Burk RD, Chen Z, van Doorslaer K, Hausen H, de Villiers EM (2010) Classification of papillomaviruses (PVs) based on 189 PV types and proposal of taxonomic amendments. Virology 401: 70-79. doi: 10.1016/j.virol.2010.02.002.
  • Bruni L, Barrionuevo-Rosas L, Albero G, Aldea M, Serrano B, Valencia S, Brotons M, Mena M, Cosano R, Muñoz J, Bosch FX, de Sanjosé S, Castellsagué X, ICO Information Centre on HPV and Cancer (HPV Information Centre) (2015) Human Papillomavirus and Related Diseases in the World. Summary Report 2015, 12-23.
  • Burk RD, Harari A, Chen Z (2013) Human papillomavirus genome variants. Virology 445: 232-243. doi: 10.1016/j.virol.2013.07.018.
  • Burroni E, Bisanzi S, Sani C, Puliti D, Carozzi F (2013) Codon 72 polymorphism of p53 and HPV type 16 E6 variants as risk factors for patients with squamous epithelial lesion of the uterine cervix.J Med Virol 85: 83-90. doi: 10.1002/jmv.23417.
  • Cornet I, Gheit T, Franceschi S, Vignat J, Burk RD, Sylla BS, Tommasino M, Clifford GM (2012) Human papillomavirus type 16 genetic variants: phylogeny and classification based on E6 and LCR. IARC HPV Variant Study Group. J Virol 86: 6855-6861. doi: 10.1128/JVI.00483-12.
  • Cornet I, Gheit T, Iannacone MR, Vignat J, Sylla BS, Del Mistro A, Franceschi S, Tommasino M, Clifford GM (2013a) HPV16 genetic variation and the development of cervical cancer worldwide. Br J Cancer 108: 240-244. doi: 10.1038/bjc.2012.508.
  • Cornet I, Gheit T, Clifford GM, Combes JD, Dalstein V, Franceschi S, Tommasino M, Clavel C (2013b) Human papillomavirus type 16 E6 variants in France and risk of viral persistence. Infect Agent Cancer 8: 4. doi: 10.1186/1750-9378-8-4.
  • Crow JM. (2012) HPV: the global burden. Nature 488: S2-S3. doi: 10.1038/488S2a.
  • de Araujo Souza PS, Sichero L, Maciag PC (2009) HPV variants and HLA polymorphisms: the role of variability on the risk of cervical cancer. Future Oncol 5: 359-370. doi: 10.2217/fon.09.8.
  • de Villiers EM, Fauquet C, Broker TR, Bernard HU, zur Hausen H (2004) Classification of papillomaviruses. Virology 324: 17-27. doi: 10.1002/(SICI)1097-0215(19961021)69:5<364: :AID-IJC2>3.0.CO;2-3.
  • Freitas LB, Chen Z, Muqui EF, Boldrini NA, Miranda AE, Spano LC, Burk RD (2014) Human papillomavirus 16 non-European variants are preferentially associated with high-grade cervical lesions. PLoS One 9: e100746. doi: 10.1371/journal.pone.0100746.
  • Garbuglia AR. (2014) Human papillomavirus in head and neck cancer. Cancers 6: 1705-1726. doi: 10.3390/cancers6031705.
  • Gheit T, Cornet I, Clifford GM, Iftner T, Munk C, Tommasino M, Kjaer SK (2011) Risks for persistence and progression by human papillomavirus type 16 variant lineages among a population-based sample of Danish women.Cancer Epidemiol Biomarkers Prev 20: 1315-1321. doi: 10.1158/1055-9965.EPI-10-1187.
  • Grodzki M, Besson G, Clavel C, Arslan A, Franceschi S, Birembaut P, Tommasino M, Zehbe I (2006) Increased risk for cervical disease progression of French women infected with the human papillomavirus type 16 E6-350G variant. Cancer Epidemiol Biomarkers Prev 15: 820-822. doi: 10.1158/1055-9965.EPI-05-0864.
  • Gudlevičienė Ž, Stumbrytė A, Juknė G, Simanavičienė V, Žvirblienė A (2015) Distribution of human papillomavirus type 16 variants in Lithuanian women with cervical cancer. Medicina (Kaunas) 51: 328-335. doi: 10.1016/j.medici.2015.11.005.
  • Han L, Maimaitiming T, Husaiyin S, Wang L, Wusainahong K, Ma C, Niyazi M (2015) Comparative study of HPV16 integration in cervical lesions between ethnicities with high and low rates of infection with high-risk HPV and the correlation between integration rate and cervical neoplasia. Exp Ther Med 10: 2169-2174. doi: 10.3892/etm.2015.2740.
  • Hildesheim A, Schiffman M, Bromley C, Wacholder S, Herrero R, Rodriguez A, Bratti MC, Sherman ME, Scarpidis U, Lin QQ, Terai M, Bromley RL, Buetow K, Apple RJ, Burk RD (2001) Human papillomavirus type 16 variants and risk of cervical cancer. J Natl Cancer Inst 93: 315-318. doi: 10.1093/jnci/93.4.315.
  • Huang L, Chao S, Lee B (2008) Integration of human papillomavirus type-16 and type-18 is a very early event in cervical carcinogenesis. J Clin Pathol 61: 627-631. doi: 10.1136/jcp.2007.052027.
  • Kulmala SM, Syrjänen SM, Gyllensten UB, Shabalova IP, Petrovichev N, Tosi P, Syrjänen KJ, Johansson BC (2006) Early integration of high copy HPV16 detectable in women with normal and low grade cervical cytology and histology. J Clin Pathol 59: 513-517. doi: 10.1136/jcp.2004.024570.
  • Lee K, Magalhaes I, Clavel C, Briolat J, Birembaut P, Tommasino M, Zehbe I (2008) Human papillomavirus 16 E6, L1, L2 and E2 gene variants in cervical lesion progression. Virus Res 131: 106-110. doi: 10.1016/j.virusres.2007.08.003.
  • Londesborough P, Ho L, Terry G, Cuzick J, Wheeler C, Singer A (1996) Human papillomavirus genotype as a predictor of persistence and development of high-grade lesions in women with minor cervical abnormalities. Int J Cancer 69: 364-368.
  • Marongiu L, Godi A, Parry JV, Beddows S (2014) Human papillomavirus type 16 long control region and E6 variants stratified by cervical disease stage. Infect Genet Evol 26: 8-13. doi: 10.1016/j.meegid.2014.05.009.
  • Muñoz N, Bosch FX, de Sanjosé S, Herrero R, Castellsagué X, Shah KV, Snijders PJ, Meijer CJ (2003) 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 348: 518-527. doi: 10.1056/NEJMoa021641.
  • Nindl I, Rindfleisch K, Lotz B, Schneider A, Dürst M (1999) Uniform distribution of HPV 16 E6 and E7 variants in patients with normal histology, cervical intra-epithelial neoplasia and cervical cancer. Int J Cancer 82: 203-207. doi: 10.1002/(SICI)1097-0215(19990719)82:2 <203::AID-IJC9>3.0.CO;2-9.
  • Ortiz-Ortiz J, Alarcón-Romero Ldel C, Jiménez-López MA, Garzón-Barrientos VH, Calleja-Macías I, Barrera-Saldaña HA, Leyva-Vázquez MA, Illades-Aguiar B (2015) Association of human papillomavirus 16 E6 variants with cervical carcinoma and precursor lesions in women from Southern Mexico. Virol J 12: 29. doi: 10.1186/s12985-015-0242-3.
  • Peitsaro P, Johansson B, Syrjänen S (2002) Integrated human papillomavirus type 16 is frequently found in cervical cancer precursors as demonstrated by a novel quantitative real-time PCR technique. J Clin Microbiol 40: 886-891. doi: 10.1128/JCM.40.3.886-891.2002.
  • Schiffman M, Rodriguez AC, Chen Z, Wacholder S, Herrero R, Hildesheim A, Desalle R, Befano B, Yu K, Safaeian M, Sherman ME, Morales J, Guillen D, Alfaro M, Hutchinson M, Solomon D, Castle PE, Burk RD (2010) A population-based prospective study of carcinogenic human papillomavirus variant lineages, viral persistence, and cervical neoplasia. Cancer Res 70: 3159-3169. doi: 10.1158/0008-5472.CAN-09-4179.
  • Shukla S, Mahata S, Shishodia G, Pande S, Verma G, Hedau S, Bhambhani S, Kumari A, Batra S, Basir SF, Das BC, Bharti AC (2014) Physical state & copy number of high risk human papillomavirus type 16 DNA in progression of cervical cancer. Indian J Med Res 139: 531-543.
  • Sichero L, Ferreira S, Trottier H, Duarte-Franco E, Ferenczy A, Franco EL, Villa LL (2007) High grade cervical lesions are caused preferentially by non-European variants of HPVs 16 and 18. Int J Cancer 120: 1763-1768. doi: 10.1002/ijc.22481.
  • Sichero L, Sobrinho JS, Villa LL (2012) Oncogenic potential diverge among human papillomavirus type 16 natural variants. Virology 432: 127-132. doi: 10.1016/j.virol.2012.06.011.
  • Szostek S, Klimek M, Zawilinska B, Kosz-Vnenchak M (2008a) Genotype-specific human papillomavirus detection in cervical smears. Acta Biochim Pol 55: 687-692.
  • Szostek S, Klimek M, Zawilinska B, Kosz-Vnenchak M (2008b) Physical state of human papillomavirus type 16 in cervical cell DNA. Folia Biol (Krakow) 56: 269-271. doi: 10.3409/fb.56_3-4.269-271.
  • Szostek S, Zawilińska B, Klimek M, Wójcik K, Koprynia M, Kosz-Vnenchak M (2011) Differentiation of an integrated and episomal HPV-16 DNA using real-time PCR in cervical specimens of women diagnosed with intraepithelial lesions and invasive cervical cancer. Ginekol Pol 82: 441-445 (in Polish).
  • Szostek S, Biesaga B, Zawilinska B, Klimek M, Kosz-Vnenchak M (2015) Physical state of human papillomavirus type 16 in cervical intraepithelial lesions and cancers determined by two different quantitative real-time PCR methods. Acta Biochim Pol 62: 923-928. doi: 10.18388/abp.2015_1161.
  • Tornesello ML, Losito S, Benincasa G, Fulciniti F, Botti G, Greggi S, Buonaguro L, Buonaguro FM (2011) Human papillomavirus (HPV) genotypes and HPV16 variants and risk of adenocarcinoma and squamous cell carcinoma of the cervix. Gynecol Oncol 121: 32-42. doi: 10.1016/j.ygyno.2010.12.005.
  • Tsakogiannis D, Papadopoulou A, Kontostathi G, Ruether IG, Kyriakopoulou Z, Dimitriou TG, Orfanoudakis G, Markoulatos P (2013) Molecular and evolutionary analysis of HPV16 E6 and E7 genes in Greek women. J Med Microbiol 62: 1688-1696. doi: 10.1099/jmm.0.055491-0.
  • van Duin M, Snijders PJ, Vossen MT, Klaassen E, Voorhorst F, Verheijen RH, Helmerhorst TJ, Meijer CJ, Walboomers JM (2000) Analysis of human papillomavirus type 16 E6 variants in relation to p53 codon 72 polymorphism genotypes in cervical carcinogenesis. J Gen Virol 81: 317-325. doi: 10.1159/000016735.
  • Vrtačnik Bokal E, Kocjan BJ, Poljak M, Bogovac Z, Jančar N (2010) Genomic variants of human papillomavirus genotypes 16, 18, and 33 in women with cervical cancer in Slovenia. J Obstet Gynaecol Res 36: 1204-1213. doi: 10.1111/j.1447-0756.2010.01316.x.
  • Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, Snijders PJ, Peto J, Meijer CJ, Munoz N (1999) Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 189: 12-19. doi: 10.1002/(SICI)1096-9896(199909)189:1<12::AID-PATH431>3.0.CO;2-F.
  • Xi LF, Kiviat NB, Hildesheim A, Galloway DA, Wheeler CM, Ho J, Koutsky LA (2006) Human papillomavirus type 16 and 18 variants: race-related distribution and persistence. J Natl Cancer Inst 98: 1045-1052. doi: 10.1093/jnci/djj297.
  • Xi LF, Koutsky LA, Hildesheim A, Galloway DA, Wheeler CM, Winer RL, Ho J, Kiviat NB (2007) Risk for high-grade cervical intraepithelial neoplasia associated with variants of human papillomavirus types 16 and 18. Cancer Epidemiol Biomarkers Prev 16: 4-10. doi: 10.1158/1055-9965.EPI-06-0670.
  • Yamada T, Manos MM, Peto J, Greer CE, Munoz N, Bosch FX, Wheeler CM (1997) Human papillomavirus type 16 sequence variation in cervical cancers: a worldwide perspective. J Virol 71: 2463-2472. 0022-538X/97/$04.0010.
  • Zehbe I, Wilander E, Delius H, Tommasino M (1998) Human papillomavirus 16 E6 variants are more prevalent in invasive cervical carcinoma than the prototype.Cancer Res 58: 829-833.
  • Zehbe I, Tachezy R, Mytilineos J, Voglino G, Mikyskova I, Delius H, Marongiu A, Gissmann L, Wilander E, Tommasino M (2001) Human papillomavirus 16 E6 polymorphisms in cervical lesions from different European populations and their correlation with human leukocyte antigen class II haplotypes. Int J Cancer 94: 711-716. doi: 10.1002/ijc.1520.
  • Zuna RE, Moore WE, Shanesmith RP, Dunn ST, Wang SS, Schiffman M, Blakey GL, Teel T (2009) Association of HPV16 E6 variants with diagnostic severity in cervical cytology samples of 354 women in a US population. Int J Cancer 125: 2609-2613. doi: 10.1002/ijc.24706.
  • zurHausen H (2002) Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer 2: 342-350. doi: 10.1038/nrc798.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.