Tumor Biology

, Volume 35, Issue 7, pp 7209–7216 | Cite as

Cyclin-dependent kinase 4 overexpression is mostly independent of gene amplification and constitutes an independent prognosticator for nasopharyngeal carcinoma

  • Tzu-Ju Chen
  • Sung-Wei Lee
  • Li-Ching Lin
  • Ching-Yih Lin
  • Kwang-Yu Chang
  • Chien-Feng Li
Research Article


Data mining in the public domain demonstrates that cyclin-dependent kinase 4 (CDK4) is highly expressed in nasopharyngeal carcinomas (NPC). Associated with cyclin-D, CDK4 phosphorylates and inactivates retinoblastoma (Rb) protein family members and mediates progression through the G1- to the S-phase of the cell cycle. Amplification and overexpression of CDK4 has been identified in various human malignancies. However, its expression and amplification has never been systemically evaluated in NPC. This study aimed to evaluate the amplification and expression status, correlation with clinicopathological features, and prognostic implications of CDK4 based on public domain dataset and in our well-defined cohort of NPC patients. The association between CDK4 transcript level and gene dosage was explored by analysis of an independent public domain dataset. We retrospectively assessed CDK4 immunoexpression in biopsies of 124 consecutive NPC patients devoid of initial distant metastasis and treated according to consistent guidelines. The results were correlated with clinicopathological features, local recurrence-free survival (LRFS), distant metastasis-free survival (DMeFS), and disease-specific survival (DSS). High levels of CDK4 protein were positively correlated with the T 3, 4 status (p = 0.024); N 2, 3 status (p < 0.001); and the American Joint Committee on Cancer stage 3, 4 (p < 0.001). Multivariate analysis suggested high CDK4 expression was an independent prognostic indicator of worse DMeFS (p = 0.001, hazard ratio (HR) = 3.226) and DSS (p = 0.037, HR = 1.838). Although CDK4 is frequently upregulated, its gene locus is very uncommonly amplified in NPC. CDK4 overexpression is mostly independent with gene amplification and represents a potential prognostic biomarker in NPC and may indicate tumor aggressiveness through cell cycle dysregulation.


Nasopharyngeal carcinoma Transcriptome CDK4 Prognosis 



Nasopharyngeal carcinoma


Epstein-Barr virus


World Health Organization


Cyclin-dependent kinase 4




Local recurrence-free survival


Distant metastasis-free survival


Disease-specific survival





This study is supported by Chi Mei Medical Center (CMFHR10303) and the Ministry of Health and Welfare (MOHW103-TD-B-111-05).

Conflicts of interest


Supplementary material

13277_2014_1884_MOESM1_ESM.doc (51 kb)
Supplementary Table 1 (DOC 51 kb)


  1. 1.
    Ng WT, Lee MC, Hung WM, et al. Clinical outcomes and patterns of failure after intensity-modulated radiotherapy for nasopharyngeal carcinoma. Int J Radiat Oncol Biol Phys. 2011;79:420–8.CrossRefPubMedGoogle Scholar
  2. 2.
    Lee AW, Sze WM, Au JS, et al. Treatment results for nasopharyngeal carcinoma in the modern era: the Hong Kong experience. Int J Radiat Oncol Biol Phys. 2005;61:1107–16.CrossRefPubMedGoogle Scholar
  3. 3.
    Leung TW, Tung SY, Sze WK, et al. Treatment results of 1070 patients with nasopharyngeal carcinoma: an analysis of survival and failure patterns. Head Neck. 2005;27:555–65.CrossRefPubMedGoogle Scholar
  4. 4.
    Burkhart DL, Sage J. Cellular mechanisms of tumour suppression by the retinoblastoma gene. Nat Rev Cancer. 2008;8:671–82.CrossRefPubMedGoogle Scholar
  5. 5.
    Benson C, Kaye S, Workman P, Garrett M, Walton M, de Bono J. Clinical anticancer drug development: targeting the cyclin-dependent kinases. Brit J Cancer. 2005;92:7–12.PubMedCentralCrossRefPubMedGoogle Scholar
  6. 6.
    Kao YC, Lee SW, Lin LC, et al. Fatty acid synthase overexpression confers an independent prognosticator and associates with radiation resistance in nasopharyngeal carcinoma. Tumour Biol. 2013;34(2):759–68.CrossRefPubMedGoogle Scholar
  7. 7.
    Win KT, Lee SW, Huang HY, et al. Nicotinamide N-methyltransferase overexpression is associated with Akt phosphorylation and indicates worse prognosis in patients with nasopharyngeal carcinoma. Tumour Biol. 2013;34(6):3923–31.CrossRefPubMedGoogle Scholar
  8. 8.
    Lan J, Tai HC, Lee SW, Chen TJ, Huang HY, Li CF. Deficiency in expression and epigenetic DNA Methylation of ASS1 gene in nasopharyngeal carcinoma: negative prognostic impact and therapeutic relevance. Tumour Biol. 2014;35(1):161–9.CrossRefPubMedGoogle Scholar
  9. 9.
    Lan J, Huang HY, Lee SW, et al. TOP2A overexpression as a poor prognostic factor in patients with nasopharyngeal carcinoma. Tumour Biol. 2014;35(1):179–87.CrossRefPubMedGoogle Scholar
  10. 10.
    Ma LJ, Lee SW, Lin LC, et al. Fibronectin overexpression is associated with latent membrane protein 1 expression and has independent prognostic value for nasopharyngeal carcinoma. Tumour Biol. 2014;35(2):1703–12.CrossRefPubMedGoogle Scholar
  11. 11.
    Hsu HP, Li CF, Lee SW, et al. Overexpression of stathmin 1 confers an independent prognostic indicator in nasopharyngeal carcinoma. Tumour Biol. 2013 Nov 12. [Epub ahead of print]Google Scholar
  12. 12.
    Budwit-Novotny DA, McCarty KS, Cox EB, et al. Immunohistochemical analyses of estrogen receptor in endometrial adenocarcinoma using a monoclonal antibody. Cancer Res. 1986;46:5419–25.PubMedGoogle Scholar
  13. 13.
    McClelland RA, Finlay P, Walker KJ, et al. Automated quantitation of immunocytochemically localized estrogen receptors in human breast cancer. Cancer Res. 1990;50:3545–50.PubMedGoogle Scholar
  14. 14.
    Masai H, Arai K. Cdc7 kinase complex: a key regulator in the initiation of DNA replication. J Cellular Physiol. 2002;190:287–96.CrossRefGoogle Scholar
  15. 15.
    Gstaiger M, Jordan R, Lim M, et al. Skp2 is oncogenic and overexpressed in human cancers. Proc Natl Acad Sci U S A. 2001;98:5043–8.PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Sutterluty H, Chatelain E, Marti A, et al. p45SKP2 promotes p27Kip1 degradation and induces S phase in quiescent cells. Nat Cell Biol. 1999;1:207–14.CrossRefPubMedGoogle Scholar
  17. 17.
    Seder CW, Hartojo W, Lin L, et al. Upregulated INHBA expression may promote cell proliferation and is associated with poor survival in lung adenocarcinoma. Neoplasia. 2009;11:388–96.PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Molven A. CDK4 (cyclin-dependent kinase 4). Atlas Genet Cytogenet Oncol Haematol. 2007;11:117–8.Google Scholar
  19. 19.
    Wang YL, Uhara H, Yamazaki Y, Nikaido T, Saida T. Immunohistochemical detection of CDK4 and p16INK4 proteins in cutaneous malignant melanoma. Brit J Dermatol. 1996;134:269–75.CrossRefGoogle Scholar
  20. 20.
    Poomsawat S, Buajeeb W, Khovidhunkit SO, Punyasingh J. Alteration in the expression of cdk4 and cdk6 proteins in oral cancer and premalignant lesions. J Oral Pathol Med: Off Publ Int Assoc Oral Pathol Am Acad Oral Pathol. 2010;39:793–9.CrossRefGoogle Scholar
  21. 21.
    Al-Aynati MM, Radulovich N, Ho J, Tsao MS. Overexpression of G1-S cyclins and cyclin-dependent kinases during multistage human pancreatic duct cell carcinogenesis. Clin Cancer Res: Off J Am Assoc Cancer Res. 2004;10:6598–605.CrossRefGoogle Scholar
  22. 22.
    Tang LH, Contractor T, Clausen R, et al. Attenuation of the retinoblastoma pathway in pancreatic neuroendocrine tumors due to increased cdk4/cdk6. Clin Cancer Res: Off J Am Assoc Cancer Res. 2012;18:4612–20.CrossRefGoogle Scholar
  23. 23.
    An HX, Beckmann MW, Reifenberger G, Bender HG, Niederacher D. Gene amplification and overexpression of CDK4 in sporadic breast carcinomas is associated with high tumor cell proliferation. Am J Pathol. 1999;154:113–8.PubMedCentralCrossRefPubMedGoogle Scholar
  24. 24.
    Rollbrocker B, Waha A, Louis DN, Wiestler OD, von Deimling A. Amplification of the cyclin-dependent kinase 4 (CDK4) gene is associated with high cdk4 protein levels in glioblastoma multiforme. Acta Neuropathol. 1996;92:70–4.CrossRefPubMedGoogle Scholar
  25. 25.
    Ghazizadeh M, Jin E, Shimizu H, et al. Role of cdk4, p16INK4, and Rb expression in the prognosis of bronchioloalveolar carcinomas. Respir Int Rev Thor Dis. 2005;72:68–73.Google Scholar
  26. 26.
    Wu A, Wu B, Guo J, et al. Elevated expression of CDK4 in lung cancer. J Transl Med. 2011;9:38.PubMedCentralCrossRefPubMedGoogle Scholar
  27. 27.
    Gast A, Scherer D, Chen B, et al. Somatic alterations in the melanoma genome: a high-resolution array-based comparative genomic hybridization study. Gene Chromosome Cancer. 2010;49:733–45.CrossRefGoogle Scholar
  28. 28.
    Muthusamy V, Hobbs C, Nogueira C, et al. Amplification of CDK4 and MDM2 in malignant melanoma. Gene Chromosome Cancer. 2006;45:447–54.CrossRefGoogle Scholar
  29. 29.
    Pilotti S, Della Torre G, Mezzelani A, et al. The expression of MDM2/CDK4 gene product in the differential diagnosis of well differentiated liposarcoma and large deep-seated lipoma. Brit J Cancer. 2000;82:1271–5.PubMedCentralCrossRefPubMedGoogle Scholar
  30. 30.
    Wikman H, Nymark P, Vayrynen A, et al. CDK4 is a probable target gene in a novel amplicon at 12q13.3-q14.1 in lung cancer. Gene Chromosome Cancer. 2005;42:193–9.CrossRefGoogle Scholar
  31. 31.
    Yu J, Deshmukh H, Payton JE, et al. Array-based comparative genomic hybridization identifies CDK4 and FOXM1 alterations as independent predictors of survival in malignant peripheral nerve sheath tumor. Clin Cancer Res: Off J Am Assoc Cancer Res. 2011;17:1924–34.CrossRefGoogle Scholar
  32. 32.
    Shimura T, Kakuda S, Ochiai Y, et al. Acquired radioresistance of human tumor cells by DNA-PK/AKT/GSK3beta-mediated cyclin D1 overexpression. Oncogene. 2010;29:4826–37.CrossRefPubMedGoogle Scholar
  33. 33.
    Shimura T, Kakuda S, Ochiai Y, Kuwahara Y, Takai Y, Fukumoto M. Targeting the AKT/GSK3beta/cyclin D1/Cdk4 survival signaling pathway for eradication of tumor radioresistance acquired by fractionated radiotherapy. Int J Radiat Oncol Biol Phys. 2011;80:540–8.CrossRefPubMedGoogle Scholar
  34. 34.
    Hagen KR, Zeng X, Lee MY, et al. Silencing CDK4 radiosensitizes breast cancer cells by promoting apoptosis. Cell Div. 2013;8:10.PubMedCentralCrossRefPubMedGoogle Scholar
  35. 35.
    de Carcer G, Perez de Castro I, Malumbres M. Targeting cell cycle kinases for cancer therapy. Curr Med Chem. 2007;14:969–85.CrossRefPubMedGoogle Scholar
  36. 36.
    Fry DW, Harvey PJ, Keller PR, et al. Specific inhibition of cyclin-dependent kinase 4/6 by PD 0332991 and associated antitumor activity in human tumor xenografts. Mol Cancer Ther. 2004;3:1427–38.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Tzu-Ju Chen
    • 1
  • Sung-Wei Lee
    • 2
  • Li-Ching Lin
    • 3
  • Ching-Yih Lin
    • 4
    • 5
  • Kwang-Yu Chang
    • 6
  • Chien-Feng Li
    • 1
    • 6
    • 7
    • 8
  1. 1.Department of PathologyChi-Mei Foundation Medical CenterTainanTaiwan
  2. 2.Department of Radiation OncologyChi-Mei Medical Center, LiouyingTainanTaiwan
  3. 3.Department of Radiation OncologyChi-Mei Medical CenterTainanTaiwan
  4. 4.Division of Gastroenterology and Hepatology, Department of Internal MedicineChi-Mei Foundation Medical CenterTainanTaiwan
  5. 5.Department of Leisure, Recreation, and Tourism ManagementSouthern Taiwan University of Science and TechnologyTainanTaiwan
  6. 6.National Institute of Cancer ResearchNational Health Research InstitutesTainanTaiwan
  7. 7.Department of BiotechnologySouthern Taiwan University of Science and TechnologyTainanTaiwan
  8. 8.Graduate Institute of Medicine, College of MedicineKaohsiung Medical UniversityKaohsiungTaiwan

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