Tumor Biology

, Volume 37, Issue 1, pp 715–721 | Cite as

Introducing differential expression of human heat shock protein 27 in hepatocellular carcinoma: moving toward identification of cancer biomarker

  • Rizma Khan
  • Nadir Naveed Siddiqui
  • Ahtesham ul Haq
  • M. Ataur Rahman
Original Article


Previously, it has to be acknowledged that overexpressed heat shock protein B27 (HSPB27) have been implicated in the etiology of wide range of human cancers. However, the molecular mechanism leading to the disease initiation to progression in liver cancer is still unknown. Present work was undertaken to investigate the differentially expressed HSPB27 in association with those damages that lead to liver cancer development. For the identification of liver cancer biomarker, samples were subjected to comparative proteomic analysis using two-dimensional gel electrophoresis (2-DE) and were further validated by Western blot and immunohistochemical analysis. After validation, in silico studies were applied to demonstrate the significantly induced phosphorylated and S-nitrosylated signals. The later included the interacting partner of HSPB27, i.e., mitogen-activated protein kinase-3 and 5 (MAPK3 and 5), ubiquitin C (UBC), v-akt murine thymoma viral oncogene homolog 1 (AKT1), mitogen-activated protein kinase 14 (MAPK14), and tumor protein p53 (TP53), which bestowed with critical capabilities, namely, apoptosis, cell cycling, stress activation, tumor suppression, cell survival, angiogenesis, proliferation, and stress resistance. Taking together, these results shed new light on the potential biomarker HSPB27 that overexpression of HSPB27 did lead to upregulation of their interacting partner that together demonstrate their possible role as a novel tumor progressive agent for the treatment of metastasis in liver cancer. HSPB27 is a promising diagnostic marker for liver cancer although further large-scale studies are required. Also, molecular profiling may help pave the road to the discovery of new therapies.


Hepatitis C virus Heat shock protein B27 Two-dimensional gel electrophoresis Western blotting 



We graciously thank Dr. N. Kabir (Panjwani Center, ICCBS, and University of Karachi- Pakistan) for providing the facility of immunohistochemistry.

The authors also thank Dr. Abid Ali (HEJ, University of Karachi) and Kamran Syed (Chemical House) for providing 2-DE facility. A part of this study was performed at the Industrial Biotechnology Department of The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, Karachi, Pakistan.

Conflicts of interest


Authors’ contributions

RK and MA. R conceived of the study; RK carried out the proteomics studies; EH collected samples and participated in prognosis analysis; NNS assisted in molecular experiments; RK drafted the manuscript; all authors have read and approved the final manuscript.

Supplementary material

13277_2015_3858_MOESM1_ESM.docx (4.8 mb)
Additional File 1 (DOCX 4963 kb)
13277_2015_3858_MOESM2_ESM.docx (25 kb)
Additional File 2 (DOCX 25 kb)
13277_2015_3858_MOESM3_ESM.docx (12 kb)
Additional File 3 (DOCX 12 kb)


  1. 1.
    Bosch FX, Ribes J, Diaz M, Cléries R. Primary liver cancer: worldwide incidence and trends. Gastroenterology. 2004;127:S5–S16.CrossRefPubMedGoogle Scholar
  2. 2.
    Bosch FX, Ribes J, Cléries R, Díaz M. Epidemiology of hepatocellular carcinoma. Clin Liver Dis. 2005;9:191–211.CrossRefPubMedGoogle Scholar
  3. 3.
    Arrigo AP. sHsp as novel regulators of programmed cell death and tumorigenicity. Pathol Biol Paris. 2000;48:280–8.PubMedGoogle Scholar
  4. 4.
    Jolly C, Morimoto RI. Re: role of the heat shock response and molecular chaperones in oncogenesis and cell death. J Natl Cancer Inst. 2001;93:239–40.Google Scholar
  5. 5.
    van de Vijver MJ, He YD, van’t Veer LJ, Dai H, Hart AA, Voskuil DW, et al. A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med. 2002;347:1999–2009.CrossRefPubMedGoogle Scholar
  6. 6.
    Ciocca DR, Calderwood SK. Heat shock proteins in cancer: diagnostic, prognostic, predictive, and treatment implications. Cell Stress Chaperones. 2005;10:86–103.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Ciocca DR, Rozados VR, Cuello-Carrión FD, Gervasoni SI, Matar P, Scharovsky OG. Heat shock proteins 25 and 70 in rodent tumors treated with doxorubicin and lovastatin. Cell Stress Chaperones. 2003;8:26–36.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Gyrd-Hansen M, Nylandsted J, Jaattela M. Heat shock protein 70 promotes cancer cell viability by safeguarding lysosomal integrity. Cell Cycle. 2004;3:1484–5.CrossRefPubMedGoogle Scholar
  9. 9.
    Concannon CG, Gorman AM, Samali A. On the role of Hsp27 in regulating apoptosis. Apoptosis. 2003;8:61–70.CrossRefPubMedGoogle Scholar
  10. 10.
    Schlesinger MJ. Heat shock proteins. J Biol Chem. 1990;265:12111–4.PubMedGoogle Scholar
  11. 11.
    Pechan PM. Heat shock proteins and cell proliferation. FEBS Lett. 1991;280:1–4.CrossRefPubMedGoogle Scholar
  12. 12.
    Lebret T, Watson RW, Fitzpatrick JM. Heat shock proteins: their role in urological tumors. J Urol. 2003;169:338–46.CrossRefPubMedGoogle Scholar
  13. 13.
    Ciocca DR, Arrigo AP, Calderwood SK. Heat shock proteins and heat shock factor 1 in carcinogenesis and tumor development: an update. Arch Toxicol. 2013;87:19–48.CrossRefPubMedGoogle Scholar
  14. 14.
    Calderwood SK, Khaleque MA, Sawyer DB, Ciocca DR. Heat shock proteins in cancer: chaperones of tumorigenesis. Trends Biochem Sci. 2006;31:164–72.CrossRefPubMedGoogle Scholar
  15. 15.
    Arrigo AP, Gibert B. HSPB27 dynamic phospho-oligomeric structure dependent interactome as cancer therapeutic target. Curr Mol Med. 2012;12:1151–63.CrossRefPubMedGoogle Scholar
  16. 16.
    Arrigo AP. Pathology-dependent effects linked to small heat shock proteins expression. Scientifica. 2012. doi: 10.6064/2012/185641.PubMedPubMedCentralGoogle Scholar
  17. 17.
    Gibert B, Eckel B, Gonin V, Goldschneider D, Fombonne J, Deux B, et al. Targeting heat shock protein 27 (HSPB27) interferes with bone metastasis and tumour formation in vivo. Br J Cancer. 2012;107:63–70.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Lemieux P, Oesterreich S, Lawrence JA, Steeg PS, Hilsenbeck SG, Harvey JM, et al. The small heat shock protein hsp27 increases invasiveness but decreases motility of breast cancer cells. Invasion Metastasis. 1997;17:113–23.PubMedGoogle Scholar
  19. 19.
    Bausero MA, Page DT, Osinaga E, Asea A. Surface expression of Hsp25 and Hsp72 differentially regulates tumor growth and metastasis. Tumour Biol. 2004;25:243–51.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Nagaraja GN, Kaur P, Asea A. Role of human and mouse HSPB27 in metastasis. Curr Mol Med. 2012;12:1142–50.CrossRefPubMedGoogle Scholar
  21. 21.
    Blackburn RV, Galoforo SS, Berns CM, Armour EP, McEachern D, Corry PM, et al. Comparison of tumor growth between Hsp25- and Hsp27- transfected murine L929 cells in nude mice. Int J Cancer. 1997;72:871–7.CrossRefPubMedGoogle Scholar
  22. 22.
    Katoh M, Koninkx J, Schumacher U. Heat shock protein expression in human tumours grown in severe combined immunodeficient mice. Cancer Lett. 2000;161:113–20.CrossRefPubMedGoogle Scholar
  23. 23.
    Hsu HS, Lin JH, Huang WC, et al. Chemoresistance of lung cancer stem like cells depends on activation of Hsp27. Cancer. 2011;117:1516–28.CrossRefPubMedGoogle Scholar
  24. 24.
    Wei L, Liu TT, Wang HH, et al. Hsp27 participates in the maintenance of breast cancer stem cells through regulation of epithelial-mesenchymal transition and nuclear factor-kappaB. Breast Cancer Res. 2011;13:R101.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Gibert B, Eckel B, Fasquelle L, Moulin M, Bouhallier F, Gonin V, et al. Knock down of heat shock protein 27 (HSPB27) induces degradation of several putative client proteins. PLoS One. 2012;7:e29719.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Arrigo AP, Gibert B. Protein interactomes of three stress inducible small heat shock proteins: HSPB27, HspB5 and HspB8. Int J Hyperth. 2013;29:409–22.CrossRefGoogle Scholar
  27. 27.
    Arrigo AP. Human small heat shock proteins: protein interactomes of homo- and hetero-oligomeric complexes: an update. FEBS Lett. 2013;587:1959–69.CrossRefPubMedGoogle Scholar
  28. 28.
    Blum H, Beier H, Gross HJ. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis. 1987;8:93–9.CrossRefGoogle Scholar
  29. 29.
    Blom N, Sicheritz-Ponten T, Gupta R, Gammeltoft S, Brunak S. Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics. 2004;4:1633–49.CrossRefPubMedGoogle Scholar
  30. 30.
    Xue Y, Liu Z, Gao X, Jin C, Wen L, Yao X, et al. GPS-SNO: computational prediction of proteins S-nitrosylation sites with a modified GPS alogrithim. PLoS One. 2008;5:e11290.CrossRefGoogle Scholar
  31. 31.
    Zhang B, Kirov S, Snoddy J. WebGestalt: an integrated system for exploring gene sets in various biological contexts. Nucleic Acids Res. 2005;33:741–8.CrossRefGoogle Scholar
  32. 32.
    Jensen LJ, Kuhn M, Stark M, Chaffron S, Creevey C, Muller J, et al. STRING 8-a global view on proteins and their functional interactions in 630 organisms. Nucleic Acids Res. 2009;37:412–6.CrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Rizma Khan
    • 1
    • 2
    • 4
  • Nadir Naveed Siddiqui
    • 3
  • Ahtesham ul Haq
    • 4
  • M. Ataur Rahman
    • 3
  1. 1.Department of BiochemistryZiauddin UniversityKarachiPakistan
  2. 2.Department of Molecular GeneticsDr. Ziauddin HospitalKarachiPakistan
  3. 3.The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE)University of KarachiKarachiPakistan
  4. 4.Department of BiochemistryUniversity of Karachi-PakistanKarachiPakistan

Personalised recommendations