Abstract
Cervical cancer is one of the most common cancers in women worldwide, and its development is related to two viral oncoproteins E6 and E7 from high-risk human papillomaviruses. Aberrant expression of E-cadherin is associated with epithelial-to-mesenchymal transition (EMT), and it is frequently seen in cervical cancer. However, the underlying mechanisms involved in E-cadherin suppression in cervical cancer are not clear. We studied the effects of human papillomavirus 16 (HPV16) E6 and E7 on E-cadherin and Cdc6 (cell division cycle 6) expression in the HCT-116 cell line. We also assessed the relationship between Cdc6 and E-cadherin expression in cells expressing HPV16 E6 and E7 proteins. The results showed that HPV16 E6 and E7 proteins reduce E-cadherin expression, and HPV16 E6-expressing cells undergo a more profound suppression of E-cadherin compared with cells expressing HPV16 E7. Our results also revealed that HPV16 E6 and E7 oncoproteins induce Cdc6 expression, whereas suppression of Cdc6 protein by short hairpin RNA restores E-cadherin expression. Induction of Cdc6 expression in HCT-116 cells was greater with E6 than with E7, a finding that was consistent with the corresponding changes in E-cadherin expression. These observations suggest that Cdc6 overexpression is an important factor for E-cadherin reduction in cells expressing HPV16 E6 and E7 proteins and may have an important role in the metastasis of HPV-associated cancers.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Pisani P, Bray F, Parkin DM . Estimates of the world‐wide prevalence of cancer for 25 sites in the adult population. Int J Cancer 2002; 97: 72–81.
Rossi A, Ciafrè S, Balsamo M, Pierimarchi P, Santoro MG . Targeting the heat shock factor 1 by RNA interference: a potent tool to enhance hyperthermochemotherapy efficacy in cervical cancer. Cancer Res 2006; 66: 7678–7685.
Cogliano V, Baan R, Straif K, Grosse Y, Secretan B, El Ghissassi F et al. Carcinogenicity of human papillomaviruses. Lancet Oncol 2005; 6: 204.
Zur Hausen H . Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer 2002; 2: 342–350.
Gagnon D, Archambault J . A high-throughput cellular assay to quantify the p53-degradation activity of E6 from different human papillomavirus types. Methods Mol Bio 2015; 1249: 111–120.
Klingelhutz AJ, Foster SA, McDougall JK . Telomerase activation by the E6 gene product of human papillomavirus type 16. Nature 1996; 380: 79–82.
Lee SS, Weiss RS, Javier RT . Binding of human virus oncoproteins to hDlg/SAP97, a mammalian homolog of the Drosophila discs large tumor suppressor protein. Proc Natl Acad Sci USA 1997; 94: 6670–6675.
Roman A, Munger K . The papillomavirus E7 proteins. Virology 2013; 445: 138–168.
Bellacchio E, Paggi MG . Understanding the targeting of the RB family proteins by viral oncoproteins to defeat their oncogenic machinery. J Cell Physiol 2013; 228: 285–291.
McLaughlin-Drubin ME, Münger K . The human papillomavirus E7 oncoprotein. Virology 2009; 384: 335–344.
Hellner K, Mar J, Fang F, Quackenbush J, Münger K . HPV16 E7 oncogene expression in normal human epithelial cells causes molecular changes indicative of an epithelial to mesenchymal transition. Virology 2009; 391: 57–63.
Gumbiner BM . Regulation of cadherin-mediated adhesion in morphogenesis. Nat Rev Mol Cell Biol 2005; 6: 622–634.
Wakita H, Yamamoto Y, Furukawa F . Aberrant suprabasal P-cadherin expression in acanthotic but not psoriatic thickened epidermis. Arch Dermatol Res 2003; 295: S71–S74.
Larue L, Bellacosa A . Epithelial–mesenchymal transition in development and cancer: role of phosphatidylinositol 3′ kinase/AKT pathways. Oncogene 2005; 24: 7443–7454.
Naito A, Iwase H, Kuzushima T, Nakamura T, Kobayashi S . Clinical significance of E‐cadherin expression in thyroid neoplasms. J Surg Oncol 2001; 76: 176–180.
Carico E, Atlante M, Bucci B, Nofroni I, Vecchione A . E-cadherin and α-catenin expression during tumor progression of cervical carcinoma. Gynecol Oncol 2001; 80: 156–161.
Dong C, Wu Y, Wang Y, Wang C, Kang T, Rychahou PG et al. Interaction with Suv39H1 is critical for Snail-mediated E-cadherin repression in breast cancer. Oncogene 2013; 32: 1351–1362.
Tseng J-C, Chen H-F, Wu K-J . A twist tale of cancer metastasis and tumor angiogenesis. Histol Histopathol 2015; 30: 1283–1294.
De Craene B, Gilbert B, Stove C, Bruyneel E, Van Roy F, Berx G . The transcription factor snail induces tumor cell invasion through modulation of the epithelial cell differentiation program. Cancer Res 2005; 65: 6237–6244.
Comijn J, Berx G, Vermassen P, Verschueren K, van Grunsven L, Bruyneel E et al. The two-handed E box binding zinc finger protein SIP1 downregulates E-cadherin and induces invasion. Mol Cell 2001; 7: 1267–1278.
Villarejo A, Cortés-Cabrera Á, Molina-Ortíz P, Portillo F, Cano A . Differential role of Snail1 and Snail2 zinc fingers in E-cadherin repression and epithelial to mesenchymal transition. J Biol Chem 2014; 289: 930–941.
Eger A, Aigner K, Sonderegger S, Dampier B, Oehler S, Schreiber M et al. DeltaEF1 is a transcriptional repressor of E-cadherin and regulates epithelial plasticity in breast cancer cells. Oncogene 2005; 24: 2375–2385.
Caberg J-HD, Hubert PM, Begon DY, Herfs MF, Roncarati PJ, Boniver JJ et al. Silencing of E7 oncogene restores functional E-cadherin expression in human papillomavirus 16-transformed keratinocytes. Carcinogenesis 2008; 29: 1441–1447.
Laurson J, Khan S, Chung R, Cross K, Raj K . Epigenetic repression of E-cadherin by human papillomavirus 16 E7 protein. Carcinogenesis 2010; 31: 918–926.
Salarini R, Sahebkar A, Mirzaei H, Jaafari M, Riahi M, Hadjati J et al. Epi-drugs and Epi-miRs: Moving beyond current cancer therapies. Current Cancer Drug Targets 2015 (e-pub ahead of print).
Mirzaei H, Yazdi F, Salehi R, Mirzaei HR . SiRNA and epigenetic aberrations in ovarian cancer. Cancer Res Ther 2016; 12: 498–508.
Yoo CB, Jones PA . Epigenetic therapy of cancer: past, present and future. Nat Rev Drug Discov 2006; 5: 37–50.
Jin B, Robertson KD . DNA methyltransferases, DNA damage repair, and cancer. Epigenet Alter Oncogen 2013; 754 p 3–29.
Hirohashi S . Inactivation of the E-cadherin-mediated cell adhesion system in human cancers. Am J Pathol 1998; 153: 333–339.
Gonzalez S, Klatt P, Delgado S, Conde E, Lopez-Rios F, Sanchez-Cespedes M et al. Oncogenic activity of Cdc6 through repression of the INK4/ARF locus. Nature 2006; 440: 702–706.
Sideridou M, Zakopoulou R, Evangelou K, Liontos M, Kotsinas A, Rampakakis E et al. Cdc6 expression represses E-cadherin transcription and activates adjacent replication origins. J Cell Biol 2011; 195: 1123–1140.
Neuwald AF, Aravind L, Spouge JL, Koonin EV . AAA+: a class of chaperone-like ATPases associated with the assembly, operation, and disassembly of protein complexes. Genome Res 1999; 9: 27–43.
Lee DG, Makhov AM, Klemm RD, Griffith JD, Bell SP . Regulation of origin recognition complex conformation and ATPase activity: differential effects of single‐stranded and double‐stranded DNA binding. EMBO J 2000; 19: 4774–4782.
Ohtani K, Tsujimoto A, Ikeda M-a, Nakamura M . Regulation of cell growth-dependent expression of mammalian CDC6 gene by the cell cycle transcription factor E2F. Oncogene 1998; 17: 1777–1785.
Yan Z, DeGregori J, Shohet R, Leone G, Stillman B, Nevins JR et al. Cdc6 is regulated by E2F and is essential for DNA replication in mammalian cells. Proc Natl Acad Sci USA 1998; 95: 3603–3608.
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.
Roger L, Jullien L, Gire V, Roux P . Gain of oncogenic function of p53 mutants regulates E-cadherin expression uncoupled from cell invasion in colon cancer cells. J Cell Sci 2010; 123: 1295–1305.
Reinhold WC, Reimers MA, Maunakea AK, Kim S, Lababidi S, Scherf U et al. Detailed DNA methylation profiles of the E-cadherin promoter in the NCI-60 cancer cells. Mol Cancer Ther 2007; 6: 391–403.
Hagemann T, Bozanovic T, Hooper S, Ljubic A, Slettenaar V, Wilson J et al. Molecular profiling of cervical cancer progression. Br J Cancer 2007; 96: 321–328.
Hamkar R, Azad TM, Mahmoodi M, Seyedirashti S, Severini A, Nategh R . Prevalence of human papillomavirus in Mazandaran province. Islamic Republic Iran 2002; 8: 805–811.
Duffy CL, Phillips SL, Klingelhutz AJ . Microarray analysis identifies differentiation-associated genes regulated by human papillomavirus type 16 E6. Virology 2003; 314: 196–205.
Watson RA, Thomas M, Banks L, Roberts S . Activity of the human papillomavirus E6 PDZ-binding motif correlates with an enhanced morphological transformation of immortalized human keratinocytes. J Cell Sci 2003; 116: 4925–4934.
D'Costa ZJ, Jolly C, Androphy EJ, Mercer A, Matthews CM, Hibma MH . Transcriptional repression of E-cadherin by human papillomavirus type 16 E6. PLoS One 2012; 7: e48954.
Martel C, Harper F, Cereghini S, Noë V, Mareel M, Cremisi C . Inactivation of retinoblastoma family proteins by SV40 T antigen results in creation of a hepatocyte growth factor/scatter factor autocrine loop associated with an epithelial-fibroblastoid conversion and invasiveness. Cell Growth Differ 1997; 8: 165–178.
Arima Y, Inoue Y, Shibata T, Hayashi H, Nagano O, Saya H et al. Rb depletion results in deregulation of E-cadherin and induction of cellular phenotypic changes that are characteristic of the epithelial-to-mesenchymal transition. Cancer Res 2008; 68: 5104–5112.
Jung JK, Arora P, Pagano JS, Jang KL . Expression of DNA methyltransferase 1 is activated by hepatitis B virus X protein via a regulatory circuit involving the p16INK4a-cyclin D1-CDK 4/6-pRb-E2F1 pathway. Cancer Res 2007; 67: 5771–5778.
Liu J, Lian Z, Han S, Waye M, Wang H, Wu M et al. Downregulation of E-cadherin by hepatitis B virus X antigen in hepatocellullar carcinoma. Oncogene 2006; 25: 1008–1017.
Arora P, Kim E-O, Jung JK, Jang KL . Hepatitis C virus core protein downregulates E-cadherin expression via activation of DNA methyltransferase 1 and 3b. Cancer Lett 2008; 261: 244–252.
Iso Y, Sawada T, Okada T, Kubota K . Loss of E‐cadherin mRNA and gain of osteopontin mRNA are useful markers for detecting early recurrence of HCV‐related hepatocellular carcinoma. J Surg Oncol 2005; 92: 304–311.
Tsai C-N, Tsai C-L, Tse K-P, Chang H-Y, Chang Y-S . The Epstein–Barr virus oncogene product, latent membrane protein 1, induces the downregulation of E-cadherin gene expression via activation of DNA methyltransferases. Proc Natl Acad Sci 2002; 99: 10084–10089.
Krishna SM, Kattoor J, Balaram P . Down regulation of adhesion protein E-cadherin in Epstein–Barr virus infected nasopharyngeal carcinomas. Cancer Biomarkers 2005; 1: 271–277.
Borlado LR, Méndez J . CDC6: from DNA replication to cell cycle checkpoints and oncogenesis. Carcinogenesis 2008; 29: 237–243.
Duursma A, Agami R . P53-dependent regulation of Cdc6 protein stability controls cellular proliferation. Mol Cell Biol 2005; 25: 6937–6947.
Murphy N, Ring M, Heffron C, King B, Killalea A, Hughes C et al. p16INK4A, CDC6, and MCM5: predictive biomarkers in cervical preinvasive neoplasia and cervical cancer. J Clin Pathol 2005; 58: 525–534.
Acknowledgements
We thank the entire staff of the Department of Virology, School of Public Health, Tehran University of Medical Sciences. This research was supported by Tehran University of Medical Sciences under research no. 93-03-27-25127.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Rights and permissions
About this article
Cite this article
Faghihloo, E., Sadeghizadeh, M., Shahmahmoodi, S. et al. Cdc6 expression is induced by HPV16 E6 and E7 oncogenes and represses E-cadherin expression. Cancer Gene Ther (2016). https://doi.org/10.1038/cgt.2016.51
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1038/cgt.2016.51
This article is cited by
-
Cyclin-dependent kinases and CDK inhibitors in virus-associated cancers
Infectious Agents and Cancer (2020)
-
Evaluating the expression level of miR-9-5p and miR-192-5p in gastrointestinal cancer: introducing novel screening biomarkers for patients
BMC Research Notes (2020)
-
Dysbiosis of Oral Microbiota and Its Effect on Epithelial-Mesenchymal Transition: a Review
SN Comprehensive Clinical Medicine (2020)
-
Evaluating the expression level of HERV-K env, np9, rec and gag in breast tissue
Infectious Agents and Cancer (2019)
-
Prevalence of herpes simplex, varicella zoster and Cytomegalovirus in tumorous and adjacent tissues of patients, suffering from colorectal cancer in Iran
Translational Medicine Communications (2019)