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Triggering of death receptor apoptotic signaling by human papillomavirus 16 E2 protein in cervical cancer cell lines is mediated by interaction with c-FLIP

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Abstract

Human papillomavirus (HPV) E2 gene disruption is one of the key features of HPV-induced cervical malignant transformation. Though it is thought to prevent progression of carcinogenesis, the pro-apoptotic function of E2 protein remains poorly understood. This study shows that expression of HPV16 E2 induces apoptosis both in HPV-positive and -negative cervical cancer cell lines and leads to hyperactivation of caspase-8 and caspase-3. Activation of these signaling factors is responsible for the observed sensitivity to apoptosis upon treatment with anti-Fas antibody or TNF-α. In addition, immunoprecipitation experiments clearly show an interaction between HPV16 E2 and c-FLIP, a key regulator of apoptotic cell death mediated by death receptor signaling. Moreover, c-FLIP and a caspase-8 inhibitor protect cells from HPV16 E2-mediated apoptosis. Overexpression of c-FLIP rescues cervical cancer cells from apoptosis induced by HPV16 E2 protein expression. The data suggest that HPV16 E2 abrogates the apoptosis-inhibitory function of c-FLIP and renders the cell hypersensitive to the Fas/FasL apoptotic signal even below threshold concentration. This suggests a novel mechanism for deregulation of cervical epithelial cell growth upon HPV-induced transformation, which is of great significance in developing therapeutic strategies for intervention of cervical carcinogenesis.

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References

  1. Bosch FX, Lorincz A, Munoz N, Meijer CJ, Shah KV (2002) The causal relation between human papillomavirus and cervical cancer. J Clin Pathol 55:244–265

    CAS  PubMed  Google Scholar 

  2. Munoz N, Bosch FX, de Sanjose S et al (2003) Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med 348:518–527

    Article  PubMed  Google Scholar 

  3. Münger K, Howley PM (2002) Human papillomavirus immortalization and transformation functions. Virus Res 89:213–228

    Article  PubMed  Google Scholar 

  4. Antson AA, Burns JE, Moroz OV, Scott DJ, Sanders CM, Bronstein IB, Dodson GG, Wilson KS, Maitland NJ (2000) Structure of the intact transactivation domain of the human papillomavirus E2 protein. Nature 403:805–809

    Article  CAS  PubMed  Google Scholar 

  5. Blachon S, Demeret C (2003) The regulatory E2 proteins of human genital papillomaviruses are pro-apoptotic. Biochimie 85:813–819

    Article  CAS  PubMed  Google Scholar 

  6. Vinokurova S, Wentzensen N, Kraus I, Klaes R, Driesch C et al (2008) Type-dependent integration frequency of human papillomavirus genomes in cervical lesions. Cancer Res 68:307–313

    Article  CAS  PubMed  Google Scholar 

  7. 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 53:201–206

    Article  CAS  PubMed  Google Scholar 

  8. Nishimura A, Ono T, Ishimoto A, Dowhanick JJ, Frizzell MA, Howley PM, Sakai H (2000) Mechanisms of human papillomavirus E2-mediated repression of viral oncogene expression and cervical cancer cell growth inhibition. J Virol 74(8):3752–3760

    Article  CAS  PubMed  Google Scholar 

  9. Desaintes C, Goyat S, Garbay S, Yaniv M, Thierry F (1999) Papillomavirus E2 induces p53-independent apoptosis in HeLa cells. Oncogene 18:4538–4545

    Article  CAS  PubMed  Google Scholar 

  10. Webster K, Parish J, Pandya M, Stern PL, Clarke AR, Gaston K (2000) The human papillomavirus (HPV) 16 E2 protein induces apoptosis in the absence of other HPV proteins and via a p53-dependent pathway. J Biol Chem 275:87–94

    Article  CAS  PubMed  Google Scholar 

  11. Demeret C, Garcia-Carranca A, Thierry F (2003) Transcription independent triggering of the extrinsic pathway of apoptosis by human papillomavirus 18 E2 protein. Oncogene 22:168–175

    Article  CAS  PubMed  Google Scholar 

  12. Shoji Y, Saegusa M, Takano Y, Ohbu M, Okayasu I (1996) Correlation of apoptosis with tumour cell differentiation, progression, and HPV infection in cervical carcinoma. J Clin Pathol 49:134–138

    Article  CAS  PubMed  Google Scholar 

  13. Aguilar-Lemarroy A, Kirchhoff S, Whitaker N, Gariglio P, zur Hausen H, Krammer PH, Rösl F (2001) Differential sensitivity of human papillomavirus type 16(+) and type 18(+) cervical carcinoma cells to CD95-mediated apoptosis. Int J Cancer 93:823–831

    Article  CAS  PubMed  Google Scholar 

  14. Krueger A, Baumann S, Krammer PH, Kirchhoff S (2001) FLICE-inhibitory proteins: regulators of death receptor-mediated apoptosis. Mol Cell Biol 21:8247–8254

    Article  CAS  PubMed  Google Scholar 

  15. Djerbi M, Screpanti V, Catrina AI et al (1999) The inhibitor of death receptor signaling, FLICE-inhibitory protein defines a new class of tumor progression factors. J Exp Med 190:1025–1032

    Article  CAS  PubMed  Google Scholar 

  16. Wang W, Wang S, Song X, Sima N, Xu X et al (2007) The relationship between c-FLIP expression and human papillomavirus E2 gene disruption in cervical carcinogenesis. Gynecol Oncol 105:571–577

    Article  CAS  PubMed  Google Scholar 

  17. Dutton A, Young LS, Murray PG (2006) The role of cellular FLICE inhibitory protein (c-FLIP) in the pathogenesis and treatment of cancer. Expert Opin Ther Targets 10:27–35

    Article  CAS  PubMed  Google Scholar 

  18. Frazer IH (2004) Prevention of cervical cancer through papillomavirus vaccination. Nat Rev Immunol 4:46–54

    Article  CAS  PubMed  Google Scholar 

  19. Zou N, Lin BY, Duan F, Lee KY, Jin G et al (2000) The hinge of the human papillomavirus type 11 E2 protein contains major determinants for nuclear localization and nuclear matrix association. J Virol 74:3761–3770

    Article  CAS  PubMed  Google Scholar 

  20. Kim KH, Seong BL (2003) Pro-apoptotic function of HBV X protein is mediated by interaction with c-FLIP and enhancement of death-inducing signal. EMBO J 22:2104–2116

    Article  CAS  PubMed  Google Scholar 

  21. Lee KH, Feig C, Tchikov V, Schickel R, Hallas C et al (2006) The role of receptor internalization in CD95 signaling. EMBO J 25:1009–1023

    Article  CAS  PubMed  Google Scholar 

  22. Medema JP, Scaffidi C, Kischkel FC, Shevchenko A, Mann M, Krammer PH, Peter ME (1997) FLICE is activated by association with the CD95 death-inducing signaling complex (DISC). EMBO J 16:2794–2804

    Article  CAS  PubMed  Google Scholar 

  23. Schweiger MR, Ottinger M, You J, Howley PM (2007) Brd4-independent transcriptional repression function of the papillomavirus e2 proteins. J Virol 81:9612–9622

    Article  CAS  PubMed  Google Scholar 

  24. Thierry F, Benotmane MA, Demeret C, Mori M, Teissier S, Desaintes C (2004) A genomic approach reveals a novel mitotic pathway in papillomavirus carcinogenesis. Cancer Res 64:895–903

    Article  CAS  PubMed  Google Scholar 

  25. Ordóñez RM, Espinosa AM, Sánchez-González DJ, Armendáriz-Borunda J, Berumen J (2004) Enhanced oncogenicity of Asian-American human papillomavirus 16 is associated with impaired E2 repression of E6/E7 oncogene transcription. J Gen Virol 85:1433–1444

    Article  PubMed  Google Scholar 

  26. Romanczuk H, Howley PM (1992) Disruption of either the E1 or the E2 regulatory gene of human papillomavirus type 16 increases viral immortalization capacity. Proc Natl Acad Sci USA 89:3159–3163

    Article  CAS  PubMed  Google Scholar 

  27. Francis DA, Schmid SI, Howley PM (2000) Repression of the integrated papillomavirus E6/E7 promoter is required for growth suppression of cervical cancer cells. J Virol 74:2679–2686

    Article  CAS  PubMed  Google Scholar 

  28. Raclaw KA, Heemers HV, Kidd EM, Dehm SM, Tindall DJ (2008) Induction of FLIP expression by androgens protects prostate cancer cells from TRAIL-mediated apoptosis. Prostate 68:1696–1706

    Article  CAS  PubMed  Google Scholar 

  29. Ullenhag GJ, Mukherjee A, Watson NF, Al-Attar AH, Scholefield JH, Durrant LG (2007) Overexpression of FLIPL is an independent marker of poor prognosis in colorectal cancer patients. Clin Cancer Res 13:5070–5075

    Article  CAS  PubMed  Google Scholar 

  30. Scaffidi C, Schmitz I, Krammer PH, Peter ME (1999) The role of c-FLIP in modulation of CD95-induced apoptosis. J Biol Chem 274:1541–1548

    Article  CAS  PubMed  Google Scholar 

  31. French LE, Tschopp J (2002) Defective death receptor signaling as a cause of tumor immune escape. Semin Cancer Biol 12:51–55

    Article  CAS  PubMed  Google Scholar 

  32. Kase H, Aoki Y, Tanaka K (2003) Fas ligand expression in cervical adenocarcinoma: relevance to lymph node metastasis and tumor progression. Gynecol Oncol 90:70–74

    Article  CAS  PubMed  Google Scholar 

  33. Konno R, Takano T, Sato S et al (2000) Serum soluble fas level as a prognostic factor in patients with gynecological malignancies. Clin Cancer Res 6:3576–3580

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 30672227; 30770913; 30628029), the “973” Program of China (No. 2009CB521800), the PhD Programs Foundation of Ministry of Education of China (No. 200804871095), the Medical Scientific Research Foundation and Natural Science Foundation of Hubei Province of China (No. JX4D10, No. 2008CDB213).

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Authors and Affiliations

  1. Cancer Biology Research Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, People’s Republic of China

    Wei Wang, Yong Fang, Yan Li, Wei Li, Li Li, Linfei Han, Shujie Liao, Zhiqiang Han, Qinglei Gao, Kezhen Li, Dongrui Deng, Li Meng, Jianfeng Zhou, Shixuan Wang & Ding Ma

  2. Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, 310006, People’s Republic of China

    Ni Sima

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  1. Wei Wang
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  2. Yong Fang
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  4. Yan Li
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  13. Li Meng
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  16. Ding Ma
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Corresponding authors

Correspondence to Shixuan Wang or Ding Ma.

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Wei Wang and Yong Fan contributed equally to this manuscript.

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Wang, W., Fang, Y., Sima, N. et al. Triggering of death receptor apoptotic signaling by human papillomavirus 16 E2 protein in cervical cancer cell lines is mediated by interaction with c-FLIP. Apoptosis 16, 55–66 (2011). https://doi.org/10.1007/s10495-010-0543-3

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  • DOI: https://doi.org/10.1007/s10495-010-0543-3

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