Tumor Biology

, Volume 35, Issue 5, pp 4247–4256 | Cite as

A cancer-related protein 14-3-3ζ is a potential tumor-associated antigen in immunodiagnosis of hepatocellular carcinoma

  • Mei Liu
  • Xinxin Liu
  • Pengfei Ren
  • Jitian Li
  • Yurong Chai
  • Su-Jun Zheng
  • Yu Chen
  • Zhong-Ping Duan
  • Ning Li
  • Jian-Ying Zhang
Research Article


Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third leading cause of cancer-related deaths worldwide. Serum alpha-fetoprotein (AFP) is the conventional biomarker currently used in clinical diagnosis of this malignancy. However, AFP is not reliable for early diagnosis, and especially the sensitivity and specificity of AFP in HCC diagnosis are not optimal. Early detection of HCC is an important issue because of the very poor prognosis and usually no more than 6 months survival after diagnosis. Therefore, there is a need for the development of more sensitive and specific methods that can supplement AFP in the early detection of this cancer. In this study, autoantibody responses to 14-3-3ζ in HCC were evaluated by enzyme-linked immunosorbent assay (ELISA), western blot, and indirect immunofluorescence assay. Immunohistochemistry (IHC) with tissue array slides was also performed to analyze protein expression of 14-3-3ζ in HCC and control tissues. The prevalence of autoantibodies against 14-3-3ζ was 16.7 % (28/168) in HCC, which was significantly higher than that in liver cirrhosis (LC), chronic hepatitis (CH), and normal human sera (NHS) (P < 0.01). The average titer of autoantibodies against 14-3-3ζ in HCC sera was higher compared to that in LC, CH, and NHS (P < 0.01). In the further study, anti-14-3-3ζ antibodies have been detected in the sera from several HCC patients with serial bleeding samples. A stronger reactive band with 14-3-3ζ in western blot can be seen in sera at 9 months before the clinical diagnosis of HCC. Our preliminary data indicate that anti-14-3-3ζ autoantibodies may be potential biomarkers for early-stage HCC screening and diagnosis.


Hepatocellular carcinoma (HCC) 14-3-3ζ Tumor-associated antigens (TAAs) Immunodiagnosis 



The authors thank Dr. Eng M. Tan (The Scripps Research Institute) for his support. This work was supported by grants (SC1CA166016) from the National Institutes of Health (NIH) and by a grant of high technical personnel training item from the Beijing Health System (2011-3-083) and You'an liver disease and AIDS funding (BJYAH-2011-047, BJYAH-2011-045). We also thank the Border Biological Research Center (BBRC) Core Facilities at The University of Texas at El Paso (UTEP) for their support, which was funded by NIH grant (5G12MD007592).


  1. 1.
    Simonetti RG, Camma C, Fiorello F, Politi F, D’Amico G, Pagliaro L. Hepatocellular carcinoma. A worldwide problem and the major risk factors. Dig Dis Sci. 1991;36:962–72.CrossRefPubMedGoogle Scholar
  2. 2.
    Blumberg BS. Hepatitis B, virus, the vaccine, and the control of primary cancer of the liver. Proc Natl Acad Sci U S A. 1997;94:7121–5.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Colombo M, Sangiovanni A. Etiology, natural history and treatment of hepatocellular carcinoma. Antivir Res. 2003;60:145–50.CrossRefPubMedGoogle Scholar
  4. 4.
    Colombo M, Donato MF. Prevention of hepatocellular carcinoma. Semin Liver Dis. 2005;25:155–61.CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang JY, Dai M, Wang X, Lu WQ, Li DS, Zhang MX, et al. A case-control study of hepatitis B and C virus infection as risk factors for hepatocellular carcinoma in Henan, China. Int J Epidemiol. 1998;27:574–8.CrossRefPubMedGoogle Scholar
  6. 6.
    Zhang JY, Wang X, Han SG, Zhuang H. A case-control study of risk factors for hepatocellular carcinoma in Henan, China. Am J Trop Med Hyg. 1998;59:947–51.CrossRefPubMedGoogle Scholar
  7. 7.
    Gardino AK, Smerdon SJ, Yaffe MB. Structural determinants of 14-3-3 binding specificities and regulation of subcellular localization of 14-3-3-ligand complexes: a comparison of the X-ray crystal structures of all human 14-3-3 isoforms. Semin Cancer Biol. 2006;16(3):173–82.CrossRefPubMedGoogle Scholar
  8. 8.
    Lee IN, Chen CH, Sheu JC, Lee HS, Huang GT, Yu CY, et al. Identification of human hepatocellular carcinoma-related biomarkers by two-dimensional difference gel electrophoresis and mass spectrometry. J Proteome Res. 2005;4(6):2062–9.CrossRefPubMedGoogle Scholar
  9. 9.
    Neal CL, Yu D. 14-3-3ζ as a prognostic marker and therapeutic target for cancer. Expert Opin Ther Targets. 2010;14(12):1343–54.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Muslin AJ, Tanner JW, Allen PM, Shaw AS. Interaction of 14-3-3 with signaling proteins is mediated by the recognition of phosphoserine. Cell. 1996;84:889–97.CrossRefPubMedGoogle Scholar
  11. 11.
    Fu H, Coburn J, Collier RJ. The eukaryotic host factor that activates exoenzyme S of Pseudomonas aeruginosa is a member of the 14-3-3 protein family. Proc Natl Acad Sci U S A. 1993;90:2320–4.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Campbell JK, Gurung R, Romero S, Speed CJ, Andrews RK, Berndt MC, et al. Activation of the 43 kDa inositol polyphosphate 5-phosphatase by 14-3-3zeta. Biochemistry. 1997;36:15363–70.CrossRefPubMedGoogle Scholar
  13. 13.
    Masters SC, Pederson KJ, Zhang L, Barbieri JT, Fu H. Interaction of 14-3-3 with a nonphosphorylated protein ligand, exoenzyme S of Pseudomonas aeruginosa. Biochemistry. 1999;38:5216–21.CrossRefPubMedGoogle Scholar
  14. 14.
    Obsil T, Obsilova V. Structural basis of 14-3-3 protein functions. Semin Cell Dev Biol. 2011;22(7):663–72.CrossRefPubMedGoogle Scholar
  15. 15.
    Freeman AK, Morrison DK. 14-3-3 proteins: diverse functions in cell proliferation and cancer progression. Semin Cell Dev Biol. 2011;22(7):681–7.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Johnson PJ, Leung N, Cheng P, Welby C, Leung WT, Lau WY, et al. ‘Hepatoma-specific’ alphafetoprotein may permit preclinical diagnosis of malignant change in patients with chronic liver disease. Br J Cancer. 1997;75:236–40.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Zhang JY, Casiano CA, Peng XX, Koziol JA, Chan EKL, Tan EM. Enhancement of antibody detection in cancer using panel of recombinant tumor-associated antigens. Cancer Epidemiol Biomark Prev. 2003;12:136–43.Google Scholar
  18. 18.
    Zhang JY, Megliorino R, Peng XX, Tan EM, Chen Y, Chan EK. Antibody detection using tumor-associated antigen mini-array in diagnosing human hepatocellular carcinoma. J Hepatol. 2007;46:107–14.CrossRefPubMedGoogle Scholar
  19. 19.
    Zhang JY, Chan EKL, Peng XX, Tan EM. A novel RNA-binding protein is an autoantigen in human hepatocellular carcinoma. J Exp Med. 1999;189:1101–10.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Himoto T, Kuriyama S, Zhang JY, Chan EK, Nishioka M, Tan EM. Significance of autoantibodies against insulin-like growth factor II mRNA-binding proteins in patients with hepatocellular carcinoma. Int J Oncol. 2005;26(2):311–7.PubMedGoogle Scholar
  21. 21.
    Ersvaer E, Zhang JY, McCormack E, Olsnes A, Anensen N, Tan EM, et al. Cyclin B1 is commonly expressed in the cytoplasm of primary human acute myelogenous leukemia cells and serves as a leukemia-associated antigen associated with autoantibody response in a subset of patients. Eur J Haematol. 2007;79(3):210–25.CrossRefPubMedGoogle Scholar
  22. 22.
    Zhang J, Wang K, Zhang J, Liu SS, Dai L, Zhang JY. Using proteomic approach to identify tumor-associated proteins as biomarkers in human esophageal squamous cell carcinoma. J Proteome Res. 2011;10:2863–72.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Chen XL, Zhou L, Yang J, Shen FK, Zhao SP, Wang YL. Hepatocellular carcinoma-associated protein markers investigated by MALDI-TOF MS. Mol Med Rep. 2010;3(4):589–96.PubMedGoogle Scholar
  24. 24.
    Goc A, Abdalla M, Al-Azayzih A, Somanath PR. Rac1 activation driven by 14-3-3ζ dimerization promotes prostate cancer cell-matrix interactions, motility and transendothelial migration. PLoS One. 2012;7(7):e40594.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Liu TA, Jan YJ, Ko BS, Chen SC, Liang SM, Hung YL, et al. Increased expression of 14-3-3β promotes tumor progression and predicts extrahepatic metastasis and worse survival in hepatocellular carcinoma. Am J Pathol. 2011;179(6):2698–708.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Ko BS, Lai IR, Chang TC, Liu TA, Chen SC, Wang J, et al. Involvement of 14-3-3γ overexpression in extrahepatic metastasis of hepatocellular carcinoma. Hum Pathol. 2011;42(1):129–35.CrossRefPubMedGoogle Scholar
  27. 27.
    Liu TA, Jan YJ, Ko BS, Liang SM, Chen SC, Wang J, et al. 14-3-3ε overexpression contributes to epithelial-mesenchymal transition of hepatocellular carcinoma. PLoS One. 2013;8(3):e57968.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Ko BS, Chang TC, Hsu C, Chen YC, Shen TL, Chen SC, et al. Overexpression of 14-3-3ε predicts tumour metastasis and poor survival in hepatocellular carcinoma. Histopathology. 2011;58(5):705–11.CrossRefPubMedGoogle Scholar
  29. 29.
    Iwata N, Yamamoto H, Sasaki S, Itoh F, Suzuki H, Kikuchi T, et al. Frequent hypermethylation of CpG islands and loss of expression of the 14-3-3 sigma gene in human hepatocellular carcinoma. Oncogene. 2000;19(46):5298–302.CrossRefPubMedGoogle Scholar
  30. 30.
    Fu WM, Zhang JF, Wang H, Tan HS, Wang WM, Chen SC, et al. Apoptosis induced by 1,3,6,7-tetrahydroxyxanthone in hepatocellular carcinoma and proteomic analysis. Apoptosis. 2012;17(8):842–51.CrossRefPubMedGoogle Scholar
  31. 31.
    Choi JE, Hur W, Jung CK, Piao LS, Lyoo K, Hong SW, et al. Silencing of 14-3-3ζ over-expression in hepatocellular carcinoma inhibits tumor growth and enhances chemosensitivity to cis-diammined dichloridoplatium. Cancer Lett. 2011;303(2):99–107.CrossRefPubMedGoogle Scholar
  32. 32.
    Chen Y, Zhou YS, Qiu SM, Wang KJ, Liu SW, Peng XX, et al. Autoantibodies to tumor-associated antigens combined with abnormal alpha-fetoprotein enhance immunodiagnosis of hepatocellular carcinoma. Cancer Lett. 2010;289:32–9.CrossRefPubMedGoogle Scholar
  33. 33.
    Shao Q, Ren P, Li Y, Peng B, Dai L, Lei N, et al. Autoantibodies against glucose-regulated protein 78 as serological diagnostic biomarkers in hepatocellular carcinoma. Int J Oncol. 2012;41(3):1061–7.PubMedPubMedCentralGoogle Scholar
  34. 34.
    Zhang JY. Mini-array of multiple tumor-associated antigens to enhance autoantibody detection for immunodiagnosis of hepatocellular carcinoma. Autoimmun Rev. 2007;6:143–8.CrossRefPubMedGoogle Scholar
  35. 35.
    Zang D, Li X, Zhang L. 14-3-3ζ Overexpression and abnormal β-catenin expression are associated with poor differentiation and progression in stage I non-small cell lung cancer. Clin Exp Med. 2010;10(4):221–8.CrossRefPubMedGoogle Scholar
  36. 36.
    Kobayashi R, Deavers M, Patenia R, Rice-Stitt T, Halbe J, Gallardo S, et al. 14-3-3 zeta protein secreted by tumor associated monocytes/macrophages from ascites of epithelial ovarian cancer patients. Cancer Immunol Immunother. 2009;58(2):247–58.CrossRefPubMedGoogle Scholar
  37. 37.
    Bajpai U, Sharma R, Kausar T, Dattagupta S, Chattopadhayay TK, Ralhan R. Clinical significance of 14-3-3 zeta in human esophageal cancer. Int J Biol Markers. 2008;23(4):231–7.CrossRefPubMedGoogle Scholar
  38. 38.
    Li Z, Zhao J, Du Y, Park HR, Sun SY, Bernal-Mizrachi L, et al. Down-regulation of 14-3-3zeta suppresses anchorage-independent growth of lung cancer cells through anoikis activation. Proc Natl Acad Sci U S A. 2008;105(1):162–7.CrossRefPubMedGoogle Scholar
  39. 39.
    Bergamaschi A, Christensen BL, Katzenellenbogen BS. Reversal of endocrine resistance in breast cancer: interrelationships among 14-3-3ζ, FOXM1, and a gene signature associated with mitosis. Breast Cancer Res. 2011;13(3):R70.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Murata T, Takayama K, Urano T, Fujimura T, Ashikari D, Obinata D, et al. 14-3-3ζ, a novel androgen-responsive gene, is upregulated in prostate cancer and promotes prostate cancer cell proliferation and survival. Clin Cancer Res. 2012;18(20):5617–27.CrossRefPubMedGoogle Scholar
  41. 41.
    He Y, Wu X, Liu X, Yan G, Xu C. LC-MS/MS analysis of ovarian cancer metastasis-related proteins using a nude mouse model: 14-3-3 zeta as a candidate biomarker. J Proteome Res. 2010;9(12):6180–90.CrossRefPubMedGoogle Scholar
  42. 42.
    Waldemarson S, Krogh M, Alaiya A, Kirik U, Schedvins K, Auer G, et al. Protein expression changes in ovarian cancer during the transition from benign to malignant. J Proteome Res. 2012;11(5):2876–89.CrossRefPubMedGoogle Scholar
  43. 43.
    Hatzipetros I, Gocze P, Koszegi T, Jaray A, Szereday L, Polgar B, et al. Investigating the clinical potential for 14-3-3 zeta protein to serve as a biomarker for epithelial ovarian cancer. J Ovarian Res. 2013;6(1):79.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Mei Liu
    • 1
    • 2
  • Xinxin Liu
    • 2
  • Pengfei Ren
    • 2
  • Jitian Li
    • 2
  • Yurong Chai
    • 2
  • Su-Jun Zheng
    • 1
  • Yu Chen
    • 1
  • Zhong-Ping Duan
    • 1
  • Ning Li
    • 1
  • Jian-Ying Zhang
    • 2
  1. 1.Beijing You’an HospitalCapital Medical UniversityBeijingChina
  2. 2.Department of Biological SciencesThe University of Texas at El PasoEl PasoUSA

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