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01-05-2012 | Original Article

Sequence variation analysis of the E2 gene of human papilloma virus type 16 in cervical lesions from women in Greece

Authors: D. Tsakogiannis, I. G. A. Ruether, Z. Kyriakopoulou, V. Pliaka, A. Theoharopoulou, V. Skordas, E. Panotopoulou, C. Nepka, P. Markoulatos

Published in: Archives of Virology | Issue 5/2012

Abstract

The E2 gene of human papilloma virus is expressed at the early stage of the viral life cycle, encoding the E2 transcription factor, and regulates the expression of E6 and E7 oncogenes. Disruption of E2 gene due to viral integration inhibits the transcriptional suppression of the HPV oncogenes, inducing cell proliferation. In the present study, a total of 22 HPV16-positive cytological specimens derived from high- and low-grade cervical intraepithelial lesions were investigated in order to identify sequence variations in the HPV16 E2 ORF. The E2 gene was amplified by PCR using external and internal overlapping sets of primers. Amplicons were cloned and sequenced. Disruption sites were detected in cervical samples diagnosed as high-grade cervical intraepithelial lesions. Moreover, sequence variations were identified in the E2 ORF and specific variations were associated with non-European variants such as African type I, African type II and Asian American. A total of three new sequence variations were identified at positions 2791, 2823 (transactivation domain) and 3361 (hinge region). Distinct phylogenetic branches were formed according to E2 analysis that characterized the different HPV16 variants. It was ascertained that non-European variants are circulating in the Greek population.

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 401:70–79PubMedCrossRef

Zur Hausen H (1996) Papillomavirus infections—a major cause of human cancers. Biochim Biophys Acta 1288:F55–F78PubMed

Bernard HU, Calleja-Macias IE, Dunn ST (2006) Genome variation of human papillomavirus types: phylogenetic and medical implications. Int J Cancer 118:1071–1076PubMedCrossRef

Yamada T, Wheeler CM, Halpern AL, Stewart ACM, Hildesheim A, Jenison SA (1995) Human Papillomavirus type 16 variant lineages in United States populations characterized by nucleotide sequence analysis of the E6, L2 and L1 coding segments. J Virol 69:7743–7753PubMed

Bosch FX, Burchell AN, Schiffman M, Giuliano AR, de Sanjose S, Bruni L, Tortolero-Luna G, Kjaer SK, Muñoz N (2008) Epidemiology and natural history of human papillomavirus infections and type-specific implications in cervical neoplasia. Vaccine 26:K1–K17PubMedCrossRef

Schiffman M, Clifford G, Buonaguro FM (2009) Classification of weakly carcinogenic human papillomavirus types: addressing the limits of epidemiology at the borderline. Infect Agent Cancer 4:8PubMedCrossRef

Sundström K, Eloranta S, Sparén P, Arnheim Dahlström L et al (2010) Prospective study of human papillomavirus (HPV) types, HPV persistence, and risk of squamous cell carcinoma of the cervix. Cancer Epidemiol Biomarkers Prev 19:2469–2478PubMedCrossRef

Castle PE, Schiffman M, Wheeler CM, Wentzensen N, Gravitt PE (2010) Human papillomavirus genotypes in cervical intraepithelial neoplasia grade 3. Cancer Epidemiol Biomarkers Prev 19:1675–1681PubMedCrossRef

Bruni L, Diaz M, Castellsagué X, Ferrer E, Bosch FX, de Sanjosé S (2010) Cervical human papillomavirus prevalence in 5 continents: meta-analysis of 1 million women with normal cytological findings. J Infect Dis 202:1789–1799PubMedCrossRef

10.

Dong SM, Kim HS, Rha SH, Sidransky D (2001) Promoter hypermethylation of multiple genes in carcinoma of the uterine cervix. Clin Cancer Res 7:1982–1986PubMed

11.

Feng Q, Balasubramanian A, Hawes SE, Toure P, Sow PS, Dem A, Dembele B, Critchlow CW, Xi L, Lu H, McIntosh MW, Young AM, Kiviat NB (2005) Detection of hypermethylated genes in women with and without cervical neoplasia. J Natl Cancer Inst 97:273–282PubMedCrossRef

12.

Smotkin D, Wettstein FO (1986) Transcription of human papillomavirus type 16 early genes in a cervical cancer and a cancer-derived cell line and identification of the E7 protein. Proc Natl Acad Sci USA 83:4680–4684PubMedCrossRef

13.

Soeda E, Ferran MC, Baker CC, McBride AA (2006) Repression of HPV16 early region transcription by the E2 protein. Virology 351:29–41PubMedCrossRef

14.

Longworth MS, Laimins LA (2004) Pathogenesis of human papillomaviruses in differentiating epithelia. Microbiol Mol Biol Rev 68:362–372PubMedCrossRef

15.

Hamid NA, Brown C, Gaston K (2009) The regulation of cell proliferation by the papillomavirus early proteins. Cell Mol Life Sci 66:1700–1717PubMedCrossRef

16.

Huang LW, Choa SL, Lee BH, Lee BH (2008) Integration of human papillomavirus type-16 and type-18 is a very early event in cervical carcinogenesis. J Clin Pathol 61:627–631PubMedCrossRef

17.

Cricca M, Morselli-Labate AM, Venturoli S et al (2007) Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions. Gynecol Oncol 106(549):557

18.

Briolat J, Dalstein V, Saunier M et al (2007) HPV prevalence, viral load and physical state of HPV-16 in cervical smears of patients with different grades of CIN. Int J Cancer 121:2198–2204PubMedCrossRef

19.

Arias-Pulido H, Peyton CL, Joste NE, Vargas H, Wheeler CM (2006) Human papillomavirus type 16 integration in cervical carcinoma in situ and in invasive cervical cancer. J Clin Microbiol 44:1755–1762PubMedCrossRef

20.

Li W, Wang W, Si M et al (2008) The physical state of HPV16 infection and its clinical significance in cancer precursor lesion and cervical carcinoma. J Cancer Res Clin Oncol 134:1355–1361PubMedCrossRef

21.

Tonon SA, Picconi MA, Bos PD, Zinovich JB, Galuppo J, Alonio LV, Teyssie AR (2001) Physical status of the E2 human papilloma virus 16 viral gene in cervical preneoplastic and neoplastic lesions. J Clin Virol 21:129–134PubMedCrossRef

22.

De Marco L, Gillio-Tos A, Bonello L, Ghisetti V, Ronco G, Merletti F (2007) Detection of human papillomavirus type 16 integration in preneoplastic cervical lesions and confirmation by DIPS-PCR and sequencing. J Clin Virol 38:7–13PubMedCrossRef

23.

Andersson S, Safari H, Mints M, Lewensohn-Fuchs I, Gyllensten U, Johansson B (2005) Type distribution, viral load and integration state of high-risk human papillomaviruses in pre-stages of cervical cancer (CIN). Br J Cancer 92:2195–2200PubMedCrossRef

24.

Hudelist G, Manavi M, Pischinger KID, Watkins-Riedel T, Singer CF, Kubista E, Czerwenka KF (2004) Physical state and expression of HPV DNA in benign and dysplastic cervical tissue: different levels of viral integration are correlated with lesion grade. Gynecol Oncol 92:873–880PubMedCrossRef

25.

Pett MR, Herdman MT, Palmer RD, Yeo GSH, Shivji MK, Stanley MA, Coleman N (2006) Selection of cervical keratinocytes containing integrated HPV16 associates with episome loss and an endogenous antiviral response. Proc Natl Acad Sci USA 103:3822–3827PubMedCrossRef

26.

Matovina M, Sabol I, Grubisić G, Gasperov NM, Grce M (2009) Identification of human papillomavirus type 16 integration sites in high-grade precancerous cervical lesions. Gynecol Oncol 113:120–127PubMedCrossRef

27.

Cheung JL, Lo KW, Cheung TH, Tang JW, Chan PK (2006) Viral load, E2 gene disruption status, and lineage of human papillomavirus type 16 infection in cervical neoplasia. J Infect Dis 194:1706–1712PubMedCrossRef

28.

Doeberitz V, Bauknecht M, Bartsch T, Zur Hauzen H (1991) Influence of chromosomal integration on glucocorticoid-regulated transcription of growth-stimulating papillomavirus genes E6 and E7 in cervical carcinoma cells. Proc Natl Acad Sci USA 18:1411–1415CrossRef

29.

Jeon S, Lambert PF (1995) Integration of human papillomavirus type 16 DNA into the human genome leads to increased stability of E6 and E7 mRNAs: implications for cervical carcinogenesis. Proc Natl Acad Sci USA 92:1654–1658PubMedCrossRef

30.

Jeon S, Allen-Hoffmann B, Lambert P (1995) Integration of human papillomavirus type 16 into the human genome correlates with a selective growth advantage of cells. J Virol 69:2989–2997PubMed

31.

Boulet G, Horvath C, Vanden Broeck D, Sahebali S, Bogers J (2007) Human papillomavirus: E6 and E7 oncogenes. Int J Biochem Cell Biol 39:2006–2011PubMedCrossRef

32.

Storey A, Greenfield I, Banks L, Pim D, Crook T, Crawford L, Stanley M (1992) Lack of immortalizing activity of a human papillomavirus type 16 variant DNA with a mutation in the E2 gene isolated from normal human cervical keratinocytes. Oncogene 7:459–465PubMed

33.

Casas I, Powell L, Klapper PE, Cleator GM (1995) New method for the extraction of viral RNA and DNA from cerebrospinal fluid for use in the polymerase chain reaction assay. J Virol Methods 53:25–36PubMedCrossRef

34.

Puranen M, Saarikoski S, Syrjänen K, Syrjänen S (1996) Polymerase chain reaction amplification of human papillomavirus DNA from archival, Papanicolaou-stained cervical smears. Acta Cytol 40:391–395PubMedCrossRef

35.

Sotlar K, Diemer D, Dethleffs A, Hack Y et al (2004) Detection and typing of human papillomavirus by e6 nested multiplex PCR. J Clin Microbiol 42:3176–3184PubMedCrossRef

36.

Collins SI, Constandinou-Williams C, Wen K, Young LS, Roberts S, Murray PG, Woodman CB (2009) Disruption of the E2 gene is a common and early event in the natural history of cervical human papillomavirus infection: a longitudinal cohort study. Cancer Res 69:3828–3832PubMedCrossRef

37.

Kulmala SM, Syrjδnen SM, Gyllensten UB et al (2006) Early integration of high copy HPV16 detectable in women with normal and low grade cervical cytology and histology. J Clin Pathol 59:513–517PubMedCrossRef

38.

Peitsaro P, Johansson B, Syrjanen 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 40:886–891PubMedCrossRef

39.

Gallo G, Bibbo M, Bagella L et al (2003) Study of viral integration of HPV-16 in young patients with LSIL. J Clin Pathol 56:532–536PubMedCrossRef

40.

Azizi N, Brazete J, Hankins C, Money D et al (2008) Canadian Women’s HIV Study Group. Influence of human papillomavirus type 16 (HPV-16) E2 polymorphism on quantification of HPV-16 episomal and integrated DNA in cervicovaginal lavages from women with cervical intraepithelial neoplasia. J Gen Virol 89:1716–1728PubMedCrossRef

41.

Giannoudis A, Duin M, Snijders PJ, Herrington CS (2001) Variation in the E2-binding domain of HPV 16 is associated with high-grade squamous intraepithelial lesions of the cervix. Br J Cancer 84:1058–1063PubMedCrossRef

42.

Jiang M, Baseman JG, Koutsky LA, Feng Q, Mao C, Kiviat NB, Xi LF (2009) Sequence variation of human papillomavirus type 16 and measurement of viral integration by quantitative PCR. J Clin Microbiol 47:521–526PubMedCrossRef

43.

Graham DA, Herrington CS (2000) HPV-16 E2 gene disruption and sequence variation in CIN 3 lesions and invasive squamous cell carcinomas of the cervix: relation to numerical chromosome abnormalities. Mol Pathol 53:201–206PubMedCrossRef

44.

Eriksson A, Herron JR, Yamada T et al (1999) Human papillomavirus type 16 variant lineages characterised by nucleotide sequence analysis of the E5 coding segment and the E2 hinge region. J Gen Virol 80:595–600PubMed

45.

Casas L, Galvan SC, Ordoñez RM, Lopez N, Guido M, Berumen J (1999) Asian-american variants of human papillomavirus type 16 have extensive mutations in the E2 gene and are highly amplified in cervical carcinomas. Int J Cancer 83:449–455PubMedCrossRef

46.

Sathish N, Abraham P, Peedicayil A, Sridharan G, Shaji RV, Chandy G (2004) E2 sequence variations of HPV 16 among patients with cervical neoplasia seen in the Indian subcontinent. Gynecol Oncol 95:363–369PubMedCrossRef

47.

Watts KJ, Thompson CH, Cossart YE, Rose BR (2002) Sequence variation and physical state of human papillomavirus type 16 cervical cancer isolates from Australia and New Caledonia. Int J Cancer 97:868–874PubMedCrossRef

48.

Swan DC, Rajeevan M, Tortolero-Luna G, Follen M, Tucker RA, Unger ER (2005) Human papillomavirus type 16 E2 and E6/E7 variants. Gynecol Oncol 96:695–700PubMedCrossRef

49.

Meissner J (1997) Sequencing errors in reference HPV clones. In: Myers G, Munger K, Sverdrup F, McBride AA, Bernard HU, Meissner J (eds) Human Papillomavirus 1997: a compilation and analysis of nucleic acid and amino acid sequences. Los Alamos National Laboratory, NM, pp 110–123

50.

Veress G, Szarka K, Dong XP, Gergely L, Pfister H (1999) Functional significance of sequence variation in the E2 gene and the long control region of human papillomavirus type 16. J Gen Virol 80:1035–1043PubMed

51.

Piccini A, Storey A, Massimi P, Banks L (1995) Mutations in the human papillomavirus type 16 E2 protein identify multiple regions of the protein involved in binding to E1. J Gen Virol 76:2909–2913PubMedCrossRef

Title: Sequence variation analysis of the E2 gene of human papilloma virus type 16 in cervical lesions from women in Greece
Authors: D. Tsakogiannis
I. G. A. Ruether
Z. Kyriakopoulou
V. Pliaka
A. Theoharopoulou
V. Skordas
E. Panotopoulou
C. Nepka
P. Markoulatos
Publication date: 01-05-2012
Publisher: Springer Vienna
Published in: Archives of Virology / Issue 5/2012
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI: https://doi.org/10.1007/s00705-012-1236-8

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