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Journal of General Virology header logo Journal of General Virology

Volume 98, Issue 9

Two common variants of human papillomavirus type 16 E6 differentially deregulate sugar metabolism and hypoxia signalling in permissive human keratinocytes No Access

Abstract

Human papillomavirus type 16 (HPV16) is responsible for most cancers attributable to HPV infection and naturally occurring variants of the HPV16 E6 oncoprotein predispose individuals to varying risk for developing cancer. Population studies by us and others have demonstrated that the common Asian–American E6 (AAE6) variant is a higher risk factor for cervical cancer than the E6 of another common variant, the European prototype (EPE6). However, a complete understanding of the molecular processes fundamental to these epidemiological findings is still lacking. Our previously published functional studies of these two E6 variants showed that AAE6 had a higher immortalization and transformation potential than EPE6. Proteomic analysis revealed markedly different protein patterns between these variants, especially with respect to key cellular metabolic enzymes. Here, we tested the Warburg effect and hypoxia signalling (hallmarks of cancer development) as plausible mechanisms underlying these observations. Lactate and glucose production were enhanced in AAE6-transduced keratinocytes, likely due to raised levels of metabolic enzymes, but independent of hypoxia-inducible factor 1 alpha (HIF-1α) activity. The HIF-1α protein level and activity were elevated by AAE6 in hypoxic conditions, leading to a hypoxia-tolerant phenotype with enhanced migratory potential. The deregulation of HIF-1α was caused by the AAE6 variant’s ability to augment mitogen-activated protein kinase/extracellular related kinase signalling. The present study reveals prominent underlying mechanisms of the AAE6’s enhanced oncogenic potential.

  • Received:
  • Accepted:
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Keyword(s): extracellular related kinase (ERK1/2) , HPV16 E6 variants , Human papillomavirus (HPV) , hypoxia , mitogen-activated protein kinase (MAPK) and warburg effect
© 2017 The Authors
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/content/journal/jgv/10.1099/jgv.0.000905
2017-09-01
2025-03-21
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References

  1. Crow JM. HPV: The global burden. Nature 2012; 488:S2–S3 [View Article][PubMed]
     [Google Scholar]
  2. Tommasino M. The human Papillomavirus family and its role in carcinogenesis. Semin Cancer Biol 2014; 26:13–21 [View Article][PubMed]
     [Google Scholar]
  3. Niccoli S, Abraham S, Richard C, Zehbe I. The Asian-American E6 variant protein of human papillomavirus 16 alone is sufficient to promote immortalization, transformation, and migration of primary human foreskin keratinocytes. J Virol 2012; 86:12384–12396 [View Article][PubMed]
     [Google Scholar]
  4. Berumen J, Ordoñez RM, Lazcano E, Salmeron J, Galvan SC et al. Asian-American variants of human Papillomavirus 16 and risk for cervical cancer: a case-control study. J Natl Cancer Inst 2001; 93:1325–1330 [View Article][PubMed]
     [Google Scholar]
  5. Sichero L, Ferreira S, Trottier H, Duarte-Franco E, Ferenczy A et al. High grade cervical lesions are caused preferentially by non-European variants of HPVs 16 and 18. Int J Cancer 2007; 120:1763–1768 [View Article][PubMed]
     [Google Scholar]
  6. Xi LF, Koutsky LA, Hildesheim A, Galloway DA, Wheeler CM et al. Risk for high-grade cervical intraepithelial neoplasia associated with variants of human Papillomavirus types 16 and 18. Cancer Epidemiol Biomarkers Prev 2007; 16:4–10 [View Article][PubMed]
     [Google Scholar]
  7. Zuna RE, Moore WE, Shanesmith RP, Dunn ST, Wang SS et al. Association of HPV16 E6 variants with diagnostic severity in cervical cytology samples of 354 women in a US population. Int J Cancer 2009; 125:2609–2613 [View Article][PubMed]
     [Google Scholar]
  8. Richard C, Lanner C, Naryzhny SN, Sherman L, Lee H et al. The immortalizing and transforming ability of two common human Papillomavirus 16 E6 variants with different prevalences in cervical cancer. Oncogene 2010; 29:3435–3445 [View Article][PubMed]
     [Google Scholar]
  9. Vander Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 2009; 324:1029–1033 [View Article][PubMed]
     [Google Scholar]
  10. Estrella V, Chen T, Lloyd M, Wojtkowiak J, Cornnell HH et al. Acidity generated by the tumor microenvironment drives local invasion. Cancer Res 2013; 73:1524–1535 [View Article][PubMed]
     [Google Scholar]
  11. Hsu PP, Sabatini DM. Cancer cell metabolism: Warburg and beyond. Cell 2008; 134:703–707 [View Article][PubMed]
     [Google Scholar]
  12. Schwickert G, Walenta S, Sundfør K, Rofstad EK, Mueller-Klieser W. Correlation of high lactate levels in human cervical cancer with incidence of metastasis. Cancer Res 1995; 55:4757–4759[PubMed]
     [Google Scholar]
  13. Walenta S, Wetterling M, Lehrke M, Schwickert G, Sundfør K et al. High lactate levels predict likelihood of metastases, tumor recurrence, and restricted patient survival in human cervical cancers. Cancer Res 2000; 60:916–921[PubMed]
     [Google Scholar]
  14. Cairns RA, Harris IS, Mak TW. Regulation of cancer cell metabolism. Nat Rev Cancer 2011; 11:85–95 [View Article][PubMed]
     [Google Scholar]
  15. Chiche J, Ilc K, Laferrière J, Trottier E, Dayan F et al. Hypoxia-inducible carbonic anhydrase IX and XII promote tumor cell growth by counteracting acidosis through the regulation of the intracellular pH. Cancer Res 2009; 69:358–368 [View Article][PubMed]
     [Google Scholar]
  16. Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003; 3:721–732 [View Article][PubMed]
     [Google Scholar]
  17. Cuninghame S, Jackson R, Zehbe I. Hypoxia-inducible factor 1 and its role in viral carcinogenesis. Virology 2014; 456–457:370–383 [View Article][PubMed]
     [Google Scholar]
  18. Birner P, Schindl M, Obermair A, Plank C, Breitenecker G et al. Overexpression of hypoxia-inducible factor 1α is a marker for an unfavorable prognosis in early-stage invasive cervical cancer. Cancer Res 2000; 60:4693–4696[PubMed]
     [Google Scholar]
  19. Kim BW, Cho H, Chung JY, Conway C, Ylaya K et al. Prognostic assessment of hypoxia and metabolic markers in cervical cancer using automated digital image analysis of immunohistochemistry. J Transl Med 2013; 11:185 [View Article][PubMed]
     [Google Scholar]
  20. Mendez LE, Manci N, Cantuaria G, Gomez-Marin O, Penalver M et al. Expression of glucose transporter-1 in cervical cancer and its precursors. Gynecol Oncol 2002; 86:138–143 [View Article][PubMed]
     [Google Scholar]
  21. Rudlowski C, Becker AJ, Schroder W, Rath W, Büttner R et al. GLUT1 messenger RNA and protein induction relates to the malignant transformation of cervical cancer. Am J Clin Pathol 2003; 120:691–698 [View Article][PubMed]
     [Google Scholar]
  22. Befani CD, Vlachostergios PJ, Hatzidaki E, Patrikidou A, Bonanou S et al. Bortezomib represses HIF-1α protein expression and nuclear accumulation by inhibiting both PI3K/Akt/TOR and MAPK pathways in prostate cancer cells. J Mol Med 2012; 90:45–54 [View Article][PubMed]
     [Google Scholar]
  23. Wong N, Ojo D, Yan J, Tang D. PKM2 contributes to cancer metabolism. Cancer Lett 2015; 356:184–191 [View Article][PubMed]
     [Google Scholar]
  24. Shestov AA, Liu X, Ser Z, Cluntun AA, Hung YP et al. Quantitative determinants of aerobic glycolysis identify flux through the enzyme GAPDH as a limiting step. Elife 2014; 3:e03342 [View Article][PubMed]
     [Google Scholar]
  25. Krishnamachary B, Berg-Dixon S, Kelly B, Agani F, Feldser D et al. Regulation of colon carcinoma cell invasion by hypoxia-inducible factor 1. Cancer Res 2003; 63:1138–1143[PubMed]
     [Google Scholar]
  26. López-Ocejo O, Viloria-Petit A, Bequet-Romero M, Mukhopadhyay D, Rak J et al. Oncogenes and tumor angiogenesis: the HPV-16 E6 oncoprotein activates the vascular endothelial growth factor (VEGF) gene promoter in a p53 independent manner. Oncogene 2000; 19:4611–4620 [View Article][PubMed]
     [Google Scholar]
  27. Wakisaka N, Kondo S, Yoshizaki T, Murono S, Furukawa M et al. Epstein-Barr virus latent membrane protein 1 induces synthesis of hypoxia-inducible factor 1α. Mol Cell Biol 2004; 24:5223–5234 [View Article][PubMed]
     [Google Scholar]
  28. Yoo YG, Oh SH, Park ES, Cho H, Lee N et al. Hepatitis B virus X protein enhances transcriptional activity of hypoxia-inducible factor-1α through activation of mitogen-activated protein kinase pathway. J Biol Chem 2003; 278:39076–39084 [View Article][PubMed]
     [Google Scholar]
  29. Chakrabarti O, Veeraraghavalu K, Tergaonkar V, Liu Y, Androphy EJ et al. Human papillomavirus type 16 E6 amino acid 83 variants enhance E6-mediated MAPK signaling and differentially regulate tumorigenesis by notch signaling and oncogenic Ras. J Virol 2004; 78:5934–5945 [View Article][PubMed]
     [Google Scholar]
  30. Dhillon AS, Hagan S, Rath O, Kolch W. MAP kinase signalling pathways in cancer. Oncogene 2007; 26:3279–3290 [View Article][PubMed]
     [Google Scholar]
  31. Murphy LO, Mackeigan JP, Blenis J. A network of immediate early gene products propagates subtle differences in mitogen-activated protein kinase signal amplitude and duration. Mol Cell Biol 2004; 24:144–153 [View Article][PubMed]
     [Google Scholar]
  32. Santarpia L, Lippman SM, El-Naggar AK. Targeting the MAPK-RAS-RAF signaling pathway in cancer therapy. Expert Opin Ther Targ 2012; 16:103–119 [View Article][PubMed]
     [Google Scholar]
  33. Togtema M, Jackson R, Richard C, Niccoli S, Zehbe I. The human Papillomavirus 16 European-T350G E6 variant can immortalize but not transform keratinocytes in the absence of E7. Virology 2015; 485:274–282 [View Article][PubMed]
     [Google Scholar]
  34. Zehbe I, Richard C, Decarlo CA, Shai A, Lambert PF et al. Human Papillomavirus 16 E6 variants differ in their dysregulation of human keratinocyte differentiation and apoptosis. Virology 2009; 383:69–77 [View Article][PubMed]
     [Google Scholar]
  35. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001; 25:402–408 [View Article][PubMed]
     [Google Scholar]
/content/journal/jgv/10.1099/jgv.0.000905
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Two common variants of human papillomavirus type 16 E6 differentially deregulate sugar metabolism and hypoxia signalling in permissive human keratinocytes
J. Gen. Virol. 98, 2310 (2017); https://doi.org/10.1099/jgv.0.000905
/content/journal/jgv/10.1099/jgv.0.000905
/content/journal/jgv/10.1099/jgv.0.000905
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