Situation of HPV16 E2 Gene Status During Radiotherapy Treatment of Cervical Carcinoma

  • Kahla, Saloua ;
  • Kochbati, Lotfi ;
  • Maalej, Mongi ;
  • Oueslati, Ridha
  • Published : 2014.03.30


Background: Human papillomavirus (HPV) integration within the E2 gene has been proposed as a critical event in cervical carcinogenesis. This study concerned whether HPV16 status and E2 gene intactness are predictive of radiation response in patients with cervical cancer. Materials and Methods: Biopsies of 44 patients with cervical cancer were collected before or after radiotherapy. The presence of HPV16 was assessed by polymerase chain reaction (PCR) using specific primers for the L1 region. E2 disruption was detected by amplifying the entire E2 gene. Results: HPV16 DNA was found in 54.5% of the clinical samples. Overall, 62.5% of the HPV16 positive tumors had integrated viral genome and 37.5% had episomal genome. There was a tendency of increase of HPV16 E2 negative tumors compared with HPV16 L1 ones in advanced stages (75% versus 20% in stage III respectively). Detection of E2 gene appeared influenced by the radiotherapy treatment, as the percentage of samples containing an intact HPV16 E2 was more frequent in pretreated patients compared to radiotherapy treated patients (66.6% versus 20%). The radiation therapy caused an eight-fold [OR= 8; CI=1.22-52.25; p=0.03] increase in the risk of HPV16 genome disruption. The integration status is influenced by the irradiation modalities, interestingly E2 disruption being found widely after radiotherapy treatment (75%) with a total fractioned dose of 50Gy. Conclusions: This study reveals that the status of the viral DNA may be used as a marker to optimize the radiation treatment.


HPV16;PCR;L1 gene;E2 gene;radiotherapy


  1. Wang CC, Lai CH, Huang HJ, et al (2010). Clinical effect of human papillomavirus genotypes in patients with cervical cancer undergoing primary radiotherapy. Int J Radiat Oncol Biol Phys, 78, 1111-20.
  2. Thierry F, Demeret C (2008). Direct activation of caspase 8 by the proapoptotic E2 protein of HPV18 independent of adaptor proteins. Cell Death Differ, 15, 1356-63.
  3. Thierry F (2009). Transcriptional regulation of the papillomavirus oncogenes by cellular and viral transcription factors in cervical carcinoma. Virology, 384, 375-9.
  4. Vozenin MC, Lord HK, Hartl D, Deutsch E (2010). Unravelling the biology of human papillomavirus (HPV) related tumours to enhance their radiosensitivity. Cancer Treat Rev, 36, 629-36.
  5. Wang L, Dai SZ, Chu HJ, Cui HF, Xu XY (2013). Integration sites and genotype distributions of human papillomavirus in cervical intraepithelial neoplasia. Asian Pac J Cancer Prev, 14, 3837-41.
  6. Yang YY, Koh LW, Tsai JH, et al (2004). Correlation of viral factors with cervical cancer in Taiwan. J Microbiol Immunol Infect, 37, 282-7.
  7. Lai CH, Chou HH, Chang CJ, et al (2013). Clinical implications of human papillomavirus genotype in cervical adenoadenosquamous carcinoma. Eur J Cancer, 49, 633-41.
  8. Gammoh N, Isaacson E, Tomaic V, et al (2009). Inhibition of HPV-16 E7 oncogenic activity by HPV-16 E2. Oncogene, 28, 2299-304.
  9. Gillison ML, Koch WM, Capone RB, et al (2000). Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst, 92, 709-20.
  10. Harima Y, Sawada S, Nagata K, Sougawa M, Ohnishi T (2001). Chromosome 6p21.2, 18q21.2 and human papilloma virus (HPV) DNA can predict prognosis of cervical cancer after radiotherapy. Int J Cancer, 96, 286-96.
  11. Le Tinier F, Reynaert N, Castelain B, et al (2012). Is adaptive intensity-modulated radiotherapy for uterine cervix carcinoma necessary? Cancer Radiother, 16, 681-7.
  12. Li H, Yang Y, Zhang R, et al (2013). Preferential sites for the integration and disruption of human papillomavirus 16 in cervical lesions. J Clin Virol, 56, 342-7.
  13. Lindel K, Beer KT, Laissue J, Greiner RH, Aebersold DM (2001). Human papillomavirus positive squamous cell carcinoma of the oropharynx: a radiosensitive subgroup of head and neck carcinoma. Cancer, 92, 805-13.<805::AID-CNCR1386>3.0.CO;2-9
  14. Lindel K, de Villiers EM, Burri P, et al (2006). Impact of viral E2-gene status on outcome after radiotherapy for patients with human papillomavirus 16-positive cancer of the uterine cervix. Int J Radiat Oncol Biol Phys, 65, 760-5.
  15. Lindel K, Burri P, Studer HU, et al (2005). Human papillomavirus status in advanced cervical cancer: predictive and prognostic significance for curative radiation treatment. Int J Gynecol Cancer, 15, 278-84.
  16. Lindel K, Rieken S, Daffinger S, et al (2012). The transcriptional regulator gene E2 of the Human Papillomavirus (HPV) 16 influences the radiosensitivity of cervical keratinocytes. Radiat Oncol, 7, 187.
  17. Micalessi IM, Boulet GA, Bogers JJ, Benoy IH, Depuydt CE (2011). High-throughput detection, genotyping and quantification of the human papillomavirus using real-time PCR. Clin Chem Lab Med, 50, 655-61.
  18. Nagai Y, Maehama T, Asato T, Kanazawa K (2000). Persistence of human papillomavirus infection after therapeutic conization for CIN 3: is it an alarm for disease recurrence? Gynecol Oncol, 79, 294-9.
  19. Parkin DM, Bray F, Ferlay J, Pisani P (2005). Global cancer statistics, 2002. CA Cancer J Clin, 55, 74-108.
  20. Santin AD, Hermonat PL, Ravaggi A, et al (1998). Radiationenhanced expression of E6/E7 transforming oncogenes of human papillomavirus-16 in human cervical carcinoma. Cancer, 83, 2346-52.<2346::AID-CNCR14>3.0.CO;2-G
  21. Schmitz M, Driesch C, Beer-Grondke K, et al (2012). Loss of gene function as a consequence of human papillomavirus DNA integration. Int J Cancer, 131, 593-602.
  22. Castellsague X, Munoz N (2003). Chapter 3: Cofactors in human papillomavirus carcinogenesis-role of parity, oral contraceptives, and tobacco smoking. J Natl Cancer Inst Monogr, 3, 20-8.
  23. Bellanger S, Tan CL, Xue YZ, Teissier S, Thierry F (2011). Tumor suppressor or oncogene? A critical role of the human papillomavirus (HPV) E2 protein in cervical cancer progression. Am J Cancer Res, 1, 373-89.
  24. Bhattacharjee B, Sengupta S (2006). HPV16 E2 gene disruption and polymorphisms of E2 and LCR: Some significant associations with cervical cancer in Indian women. Gynecol Oncol, 100, 372-8.
  25. But-Hadzic J, Jenko K, Poljak M, et al (2011). Sinonasal inverted papilloma associated with squamous cell carcinoma. Radiol Oncol, 45, 267-72.
  26. Castle PE, Lorincz AT, Mielzynska-Lohnas I, et al (2002). Results of human papillomavirus DNA testing with the hybrid capture 2 assay are reproducible. J Clin Microbiol, 40, 1088-90.
  27. Cricca M, Venturoli S, Leo E, et al (2009). Molecular analysis of HPV 16 E6I/E6II spliced mRNAs and correlation with the viral physical state and the grade of the cervical lesion. J Med Virol, 81, 1276-82.
  28. Delgado FG, Martinez E, Cespedes MA, et al (2009). Increase of human papillomavirus-16 E7-specific T helper type 1 response in peripheral blood of cervical cancer patients after radiotherapy. Immunology, 126, 523-34.
  29. Desaintes C, Goyat S, Garbay S, Yaniv M, Thierry F (1999). Papillomavirus E2 induces p53-independent apoptosis in HeLa cells. Oncogene, 18, 4538-45.
  30. Gammoh N, Grm HS, Massimi P, Banks L (2006). Regulation of human papillomavirus type 16 E7 activity through direct protein interaction with the E2 transcriptional activator. J Virol, 80, 1787-97.
  31. Alazawi W, Pett M, Strauss S, et al (2004). Genomic imbalances in 70 snap-frozen cervical squamous intraepithelial lesions: associations with lesion grade, state of the HPV16 E2 gene and clinical outcome. Br J Cancer, 91, 2063-70.

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