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Chemokine Expression Profiles of Human Hepatoma Cell Lines Mediated by Hepatitis B Virus X Protein

  • Original Article
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Pathology & Oncology Research

Abstract

The hepatitis B virus X protein (HBx), which is encoded by hepatitis B virus (HBV), plays crucial roles in the tumorigenesis of HBV associated hepatocellular carcinoma (HCC). Recent studies suggest that the HBx is involved in regulation of host immune cytokines and chemokines in HBV-associated HCC patients. However, effects of the HBx on autocrine chemokine expression profiles of hepatoma cells, which were shown in modulation of tumor-immune cell interactions, have not been investigated comprehensively. In the present study, human hepatoma cell lines SMMC-7721 and HepG2 were transfected with HBx-expressing plasmid. Human chemokine antibody array 1 (RayBio®), which simultaneously detects 38 chemokine factors, was used to determine chemokine expression profiles. Real-time polymerase chain reaction (real-time PCR) was used to further confirm the differential expression of chemokines. Chemokine antibody array revealed that all 38 chomekines were found to be expressed by SMMC-7721 and HepG2 cell lines. Interleukin-8 (IL-8) was obviously up-regulated, and epithelial neutrophil-activating protein 78 (ENA78), eosinophil chemotactic protein-1 (Eotaxin-1), monocyte chemotactic protein-1 (MCP-1), MCP-2, MCP-3 and macrophage inflammatory protein-3β (MIP-3β) were significantly declined in both cell lines following transfection of HBx-expressing plasmid. Other chemokines showed little or no significant changes. HBx-induced differential chemokine expression levels were validated by real-time PCR. Hierarchical cluster analysis identified a distinction of chomekine expression profiles between HBX-expressing hepatoma cell lines and controls. Our findings provide new evidence that HBx is able to selectively regulate chomekines in hepatoma cells that may be involved in the regulation of tumor-immune cell interactions.

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References

  1. Bosch FX, Ribes J, Diaz M, Cleries R (2004) Primary liver cancer: worldwide incidence and trends. Gastroenterology 127:S5–S16

    Article  PubMed  Google Scholar 

  2. Chen L, Zhao H, Yang X, Gao JY, Cheng J (2014) HBsAg-negative hepatitis B virus infection and hepatocellular carcinoma. Discov Med 18:189–193

    PubMed  Google Scholar 

  3. Andrisani OM (2013) Deregulation of epigenetic mechanisms by the hepatitis B virus X protein in hepatocarcinogenesis. Viruses 5:858–872

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Almajhdi FN, Al-Qudari AY, Hussain Z (2013) Differential expression of transforming growth factor-β1 and HBx enhances hepatitis B virus replication and augments host immune cytokines and chemokines. Ann Hepatol 12:408–415

    CAS  PubMed  Google Scholar 

  5. Cho HK, Kim SY, Seong JK, Cheong J (2014) Hepatitis B virus X increases immune cell recruitment by induction of chemokine SDF-1. FEBS Lett 588:733–739

    Article  CAS  PubMed  Google Scholar 

  6. Johrer K, Pleyer L, Olivier A, Maizner E, Zelle-Rieser C, Greil R (2008) Tumour-immune cell interactions modulated by chemokines. Expert Opin Biol Ther 8:269–290

    Article  PubMed  Google Scholar 

  7. O’Hayre M, Salanga CL, Handel TM, Allen SJ (2008) Chemokines and cancer: migration, intracellular signalling and intercellular communication in the microenvironment. Biochem J 409:635–649

    Article  PubMed  Google Scholar 

  8. López-Lago MA, Posner S, Thodima VJ, Molina AM, Motzer RJ, Chaganti RS (2013) Neutrophil chemokines secreted by tumor cells mount a lung antimetastatic response during renal cell carcinoma progression. Oncogene 32:1752–1760

    Article  PubMed  PubMed Central  Google Scholar 

  9. Roth MD, Harui A (2015) Human tumor infiltrating lymphocytes cooperatively regulate prostate tumor growth in a humanized mouse model. J Immunother Cancer 3:12

    Article  PubMed  PubMed Central  Google Scholar 

  10. Verbeke H, Geboes K, Van Damme J, Struyf S (2012) The role of CXC chemokines in the transition of chronic inflammation to esophageal and gastric cancer. Biochim Biophys Acta 1825:117–129

    CAS  PubMed  Google Scholar 

  11. Ben-Baruch A (2006) The multifaceted roles of chemokines in malignancy. Cancer Metastasis Rev 25:357–371

    Article  CAS  PubMed  Google Scholar 

  12. Park MH, Lee JS, Yoon JH (2012) High expression of CX3CL1 by tumor cells correlates with a good prognosis and increased tumor-infiltrating CD8+ T cells, natural killer cells, and dendritic cells in breast carcinoma. J Surg Oncol 106:386–392

    Article  CAS  PubMed  Google Scholar 

  13. Hirano S, Iwashita Y, Sasaki A, Kai S, Ohta M, Kitano S (2007) Increased mRNA expression of chemokines in hepatocellular carcinoma with tumor-infiltrating lymphocytes. J Gastroenterol Hepatol 22:690–696

    CAS  PubMed  Google Scholar 

  14. Pollicino T, Bellinghieri L, Restuccia A, Raffa G, Musolino C, Alibrandi A, et al. (2013) Hepatitis B virus (HBV) induces the expression of interleukin-8 that in turn reduces HBV sensitivity to interferon-alpha. Virology 444:317–328

    Article  CAS  PubMed  Google Scholar 

  15. Chen HY, Tang NH, Li XJ, Zhang SJ, Chen ZX, Wang XZ (2004) Transfection and expression of hepatitis B virus x gene and its effect on apoptosis in HL-7702 cells. World J Gastroenterol 10:959–964

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Luo MX, Wong SH, Chan MT, Yu L, Yu SS, Wu F, et al. (2015) Autophagy mediates HBx-induced nuclear factor-κB activation and release of IL-6, IL-8, and CXCL2 in hepatocytes. J Cell Physiol 230:2382–2389

    Article  CAS  PubMed  Google Scholar 

  17. Sun LF, Shi C, Yuan L, Sun Y, Yao XX, Ma JW, et al. (2013) Expression of cytokines in mouse hepatitis B virus X gene-transfected model. J Huazhong Univ Sci Technolog Med Sci 33:172–177

    Article  CAS  PubMed  Google Scholar 

  18. Han J, Yoo HY, Choi BH, Rho HM (2000) Selective transcriptional regulations in the human liver cell by hepatitis B viral X protein. Biochem Biophys Res Commun 272:525–530

    Article  CAS  PubMed  Google Scholar 

  19. Miyagawa T, Sugaya M, Okada A, Asano Y, Kadono T, Sato S (2014) Neutrophil-rich anaplastic large cell lymphoma expressing interleukin-8. Acta Derm Venereol 94:741–742

    Article  PubMed  Google Scholar 

  20. Cougot D, Wu Y, Cairo S, Caramel J, Renard CA, Lévy L, et al. (2007) The hepatitis B virus X protein functionally interacts with CREB-binding protein/p300 in the regulation of CREB-mediated transcription. J Biol Chem 282:4277–4287

    Article  CAS  PubMed  Google Scholar 

  21. Ehling J, Tacke F (2015) Role of chemokine pathways in hepatobiliary cancer. Cancer Lett. doi:10.1016/j.canlet.2015.06.017

  22. Li XP, Yang XY, Biskup E, Zhou J, Li HL, Wu YF, et al (2015) Co-expression of CXCL8 and HIF-1α is associated with metastasis and poor prognosis in hepatocellular carcinoma. Oncotarget 6:22880–22889

  23. Perez LE, Parquet N, Meads M, Anasetti C, Dalton W (2010) Bortezomib restores stroma-mediated APO2L/TRAIL apoptosis resistance in multiple myeloma. Eur J Haematol 84:212–222

    Article  CAS  PubMed  Google Scholar 

  24. Taub DD, Proost P, Murphy WJ, Anver M, Longo DL, van Damme J, et al. (1995) Monocyte chemotactic protein-1 (MCP-1), -2, and -3 are chemotactic for human T lymphocytes. J Clin Invest 95:1370–1376

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Wang Q, Shu C, Su J, Li X (2015) A crosstalk triggered by hypoxia and maintained by MCP-1/miR-98/IL-6/p38 regulatory loop between human aortic smooth muscle cells and macrophages leads to aortic smooth muscle cells apoptosis via Stat1 activation. Int J Clin Exp Pathol 8:2670–2679

    PubMed  PubMed Central  Google Scholar 

  26. Luo KQ, Shi YN, Peng JC (2014) The effect of chemokine CC motif ligand 19 on the proliferation and migration of hepatocellular carcinoma. Tumour Biol 35:12575–12581

    Article  CAS  PubMed  Google Scholar 

  27. Lu J, Ma J, Cai W, Wangpu X, Feng H, Zhao J, et al. (2015) CC motif chemokine ligand 19 suppressed colorectal cancer in vivo accompanied by an increase in IL-12 and IFN-γ. Biomed Pharmacother 69:374–379

    Article  CAS  PubMed  Google Scholar 

  28. Lotfi R, Lee JJ, Lotze MTJ (2007) Eosinophilic granulocytes and damage-associated molecular pattern molecules (DAMPs): role in the inflammatory response within tumors. J Immunother 30:16–28

    Article  CAS  PubMed  Google Scholar 

  29. Arenberg DA, Keane MP, DiGiovine B, Kunkel SL, Morris SB, Xue YY, et al. (1998) Epithelial-neutrophil activating peptide (ENA-78) is an important angiogenic factor in non-small cell lung cancer. J Clin Invest 102:465–472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Lim JB, Chung HW (2015) Serum ENA78/CXCL5, SDF-1/CXCL12, and their combinations as potential biomarkers for prediction of the presence and distant metastasis of primary gastric cancer. Cytokine 7:16–22

    Article  Google Scholar 

  31. Zhu F, Liu P, Li J, Zhang Y (2014) Eotaxin-1 promotes prostate cancer cell invasion via activation of the CCR3-ERK pathway and upregulation of MMP-3 expression. Oncol Rep 31:2049–2054

    CAS  PubMed  Google Scholar 

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Acknowledgments

The authors thank Professor You-Hua Xie (The Key Laboratory of Medical Molecular Virology, MOE & MOH, Fudan University) for pcDNA3.1-Flag-HBx plasmid and pcDNA3.1 plasmid.

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Authors and Affiliations

  1. Department of Pathology, Affiliated Hospital of Taishan Medical University, Taian, 271000, China

    Di-Yi Wang

  2. Department of Pathology, Huashan Hospital, Fudan University, Shanghai, 200040, China

    Li-Ping Zou

  3. Department of Pathology, Shanghai Medical College, Fudan University, Shanghai, 200032, China

    Xiao-Jia Liu, Hong-Guang Zhu & Rong Zhu

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  1. Di-Yi Wang
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Correspondence to Di-Yi Wang.

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Wang, DY., Zou, LP., Liu, XJ. et al. Chemokine Expression Profiles of Human Hepatoma Cell Lines Mediated by Hepatitis B Virus X Protein. Pathol. Oncol. Res. 22, 393–399 (2016). https://doi.org/10.1007/s12253-015-0014-9

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  • DOI: https://doi.org/10.1007/s12253-015-0014-9

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