HDAC2-mediated upregulation of IL-6 triggers the migration of osteosarcoma cells

Abstract

Understanding the mechanisms of advanced metastasis in osteosarcoma (OS) cell is important for the targeted treatment and drug development. Emerging evidence shows that epigenetic factors such as histone deacetylases (HDACs) are involved in the progression and chemoresistance of OS, while their roles are largely unknown. Our data showed that the expression of HDAC2, while not HDAC1, 4, or 8, was significantly increased in OS cells and tissues. OS patients with increased expression of HDAC2 showed reduced overall survival with p value of 0.0001. Targeted inhibition of HDAC2 suppressed the in vitro migration and invasion of OS cells. Our data showed that the inhibition of HDAC2 can decrease the expression and transcription of interleukin-6 (IL-6) in OS cells. Overexpression of IL-6 can reverse si-HDAC2-induced suppression of cell migration. Mechanistical studies showed that inhibition of HDAC2 decreased the phosphorylation and nuclear accumulation of p65, the key factor of NF-κB complex responsible for the transcription of IL-6. This was due to that HDAC2 can activate the transcription of IKK-β in OS cells. Collectively, our data showed that HDAC2-activated NF-κB can increase the expression of IL-6 in OS cells, which resulted in the promotion of cell migration. It suggested that targeted inhibition of HDAC2/NF-κB/IL-6 might be a potential approach for OS therapy.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. Al-Romaih K, Somers GR, Bayani J, Hughes S, Prasad M, Cutz JC, et al. Modulation by decitabine of gene expression and growth of osteosarcoma U2OS cells in vitro and in xenografts: identification of apoptotic genes as targets for demethylation. Cancer Cell Int. 2007;7:14.

    PubMed  PubMed Central  Google Scholar 

  2. Bennani-Baiti IM, Machado I, Llombart-Bosch A, Kovar H. Lysine-specific demethylase 1 (LSD1/KDM1A/AOF2/BHC110) is expressed and is an epigenetic drug target in chondrosarcoma, Ewing's sarcoma, osteosarcoma, and rhabdomyosarcoma. Hum Pathol. 2012;43:1300–7.

    CAS  PubMed  Google Scholar 

  3. Brasier AR. The nuclear factor-kappaB-interleukin-6 signalling pathway mediating vascular inflammation. Cardiovasc Res. 2010;86:211–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Burgess M, Tawbi H. Immunotherapeutic approaches to sarcoma. Curr Treat Options in Oncol. 2015;16:26.

    Google Scholar 

  5. Chang CC, Lin BR, Chen ST, Hsieh TH, Li YJ, Kuo MY. HDAC2 promotes cell migration/invasion abilities through HIF-1alpha stabilization in human oral squamous cell carcinoma. J Oral Pathol Med. 2011;40:567–75.

    CAS  PubMed  Google Scholar 

  6. Chen Y, Wang H, Yoon SO, Xu X, Hottiger MO, Svaren J, et al. HDAC-mediated deacetylation of NF-kappaB is critical for Schwann cell myelination. Nat Neurosci. 2011;14:437–41.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Cheng DD, Yang QC, Zhang ZC, Yang CX, Liu YW. Antitumor activity of histone deacetylase inhibitor trichostatin A in osteosarcoma cells. Asian Pac J Cancer Prev. 2012;13:1395–9.

    PubMed  Google Scholar 

  8. Choi SW, Gatza E, Hou GQ, Sun YP, Whitfield J, Song YH, et al. Histone deacetylase inhibition regulates inflammation and enhances Tregs after allogeneic hematopoietic cell transplantation in humans. Blood. 2015;125:815–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Daw NC, Chou AJ, Jaffe N, Rao BN, Billups CA, Rodriguez-Galindo C, et al. Recurrent osteosarcoma with a single pulmonary metastasis: a multi-institutional review. Br J Cancer. 2015;112:278–82.

    CAS  Google Scholar 

  10. Deng Z, Liu X, Jin J, Xu H, Gao Q, Wang Y, et al. Histone deacetylase inhibitor trichostatin A promotes the apoptosis of osteosarcoma cells through p53 signaling pathway activation. Int J Biol Sci. 2016;12:1298–308.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Fritsche P, Seidler B, Schuler S, Schnieke A, Gottlicher M, Schmid RM, et al. HDAC2 mediates therapeutic resistance of pancreatic cancer cells via the BH3-only protein NOXA. Gut. 2009;58:1399–409.

    CAS  PubMed  Google Scholar 

  12. Heinrich PC, Behrmann I, Haan S, Hermanns HM, Muller-Newen G, Schaper F. Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochem J. 2003;374:1–20.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Hunter CA, Jones SA. IL-6 as a keystone cytokine in health and disease. Nat Immunol. 2015;16:448–57.

    CAS  PubMed  Google Scholar 

  14. Itoh H, Kadomatsu T, Tanoue H, Yugami M, Miyata K, Endo M, et al. TET2-dependent IL-6 induction mediated by the tumor microenvironment promotes tumor metastasis in osteosarcoma. Oncogene. 2018;37:2903–20.

    CAS  PubMed  Google Scholar 

  15. Kesanakurti D, Chetty C, Dinh DH, Gujrati M, Rao JS. Role of MMP-2 in the regulation of IL-6/Stat3 survival signaling via interaction with alpha5beta1 integrin in glioma. Oncogene. 2013;32:327–40.

  16. Kumar P, Gogulamudi VR, Periasamy R, Raghavaraju G, Subramanian U, Pandey KN. Inhibition of HDAC enhances STAT acetylation, blocks NF-kappaB, and suppresses the renal inflammation and fibrosis in Npr1 haplotype male mice. Am J Physiol Ren Physiol. 2017;313:F781–95.

    CAS  Google Scholar 

  17. Li B, Ye Z. Epigenetic alterations in osteosarcoma: promising targets. Mol Biol Rep. 2014;41:3303–15.

    CAS  PubMed  Google Scholar 

  18. Li L, Mei DT, Zeng Y. HDAC2 promotes the migration and invasion of non-small cell lung cancer cells via upregulation of fibronectin. Biomed Pharmacother. 2016;84:284–90.

    CAS  PubMed  Google Scholar 

  19. Mook OR, Frederiks WM, Van Noorden CJ. The role of gelatinases in colorectal cancer progression and metastasis. Biochim Biophys Acta. 2004;1705:69–89.

    CAS  Google Scholar 

  20. Mu X, Brynien D, Weiss KR. The HDAC inhibitor vorinostat diminishes the in vitro metastatic behavior of osteosarcoma cells. Biomed Res Int. 2015;2015:290368.

    PubMed  PubMed Central  Google Scholar 

  21. Murahari S, Jalkanen AL, Kulp SK, Chen CS, Modiano JF, London CA, et al. Sensitivity of osteosarcoma cells to HDAC inhibitor AR-42 mediated apoptosis. BMC Cancer. 2017;17:67.

    PubMed  PubMed Central  Google Scholar 

  22. Mutsaers AJ, Walkley CR. Cells of origin in osteosarcoma: mesenchymal stem cells or osteoblast committed cells? Bone. 2014;62:56–63.

    PubMed  Google Scholar 

  23. Mutze K, Langer R, Becker K, Ott K, Novotny A, Luber B, et al. Histone deacetylase (HDAC) 1 and 2 expression and chemotherapy in gastric cancer. Ann Surg Oncol. 2010;17:3336–343.

    PubMed  Google Scholar 

  24. Nural-Guvener H, Zakharova L, Feehery L, Sljukic S, Gaballa M. Anti-fibrotic effects of class I HDAC inhibitor, mocetinostat is associated with IL-6/Stat3 signaling in ischemic heart failure. Int J Mol Sci. 2015;16:11482–99.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Pasic I, Shlien A, Durbin AD, Stavropoulos DJ, Baskin B, Ray PN, et al. Recurrent focal copy-number changes and loss of heterozygosity implicate two noncoding RNAs and one tumor suppressor gene at chromosome 3q13.31 in osteosarcoma. Cancer Res. 2010;70:160–71.

    CAS  PubMed  Google Scholar 

  26. Pittayapruek P, Meephansan J, Prapapan O, Komine M, Ohtsuki M. Role of matrix metalloproteinases in photoaging and photocarcinogenesis. Int J Mol Sci. 2016;17.

    PubMed Central  Google Scholar 

  27. Ram Kumar RM, Boro A, Fuchs B. Involvement and clinical aspects of microRNA in osteosarcoma. Int J Mol Sci. 2016;17.

    PubMed Central  Google Scholar 

  28. Raymond AK, Jaffe N. Osteosarcoma multidisciplinary approach to the management from the pathologist's perspective. Pediatr Adolescent Osteosarcoma. 2009;152:63–84.

    Google Scholar 

  29. Roh MS, Kim CW, Park BS, Kim GC, Jeong JH, Kwon HC, et al. Mechanism of histone deacetylase inhibitor trichostatin A induced apoptosis in human osteosarcoma cells. Apoptosis. 2004;9:583–9.

    CAS  PubMed  Google Scholar 

  30. Ropero S, Esteller M. The role of histone deacetylases (HDACs) in human cancer. Mol Oncol. 2007;1:19–25.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Roy SS, Gonugunta VK, Bandyopadhyay A, Rao MK, Goodall GJ, Sun LZ, et al. Significance of PELP1/HDAC2/miR-200 regulatory network in EMT and metastasis of breast cancer. Oncogene. 2014;33:3707–16.

    CAS  Google Scholar 

  32. Schafer C, Goder A, Beyer M, Kiweler N, Mahendrarajah N, Rauch A, et al. Class I histone deacetylases regulate p53/NF-kappaB crosstalk in cancer cells. Cell Signal. 2017;29:218–25.

    PubMed  Google Scholar 

  33. Schuetze KB, McKinsey TA, Long CS. Targeting cardiac fibroblasts to treat fibrosis of the heart: focus on HDACs. J Mol Cell Cardiol. 2014;70:100–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Siclari VA, Qin L. Targeting the osteosarcoma cancer stem cell. J Orthop Surg Res. 2010;5:78.

    PubMed  PubMed Central  Google Scholar 

  35. Tsuchiya H, Kanazawa Y, Abdel-Wanis ME, Asada N, Abe S, Isu K, et al. Effect of timing of pulmonary metastases identification on prognosis of patients with osteosarcoma: the Japanese musculoskeletal oncology group study. J Clin Oncol. 2002;20:3470–7.

    PubMed  Google Scholar 

  36. Tu B, Du L, Fan QM, Tang Z, Tang TT. STAT3 activation by IL-6 from mesenchymal stem cells promotes the proliferation and metastasis of osteosarcoma. Cancer Lett. 2012;325:80–8.

    CAS  PubMed  Google Scholar 

  37. Vasaikar SV, Straub P, Wang J, Zhang B. LinkedOmics: analyzing multi-omics data within and across 32 cancer types. Nucleic Acids Res. 2018;46:D956–63.

    CAS  PubMed  Google Scholar 

  38. Wagner T, Kiweler N, Wolff K, Knauer SK, Brandl A, Hemmerich P, et al. Sumoylation of HDAC2 promotes NF-kappaB-dependent gene expression. Oncotarget. 2015;6:7123–35.

    PubMed  PubMed Central  Google Scholar 

  39. Watanabe K, Okamoto K, Yonehara S. Sensitization of osteosarcoma cells to death receptor-mediated apoptosis by HDAC inhibitors through downregulation of cellular FLIP. Cell Death Differ. 2005;12:10–8.

    CAS  PubMed  Google Scholar 

  40. Weichert, W. HDAC expression and clinical prognosis in human malignancies. Cancer Lett. 2009;280:168–76.

    CAS  PubMed  Google Scholar 

  41. Wittenburg LA, Bisson L, Rose BJ, Korch C, Thamm DH. The histone deacetylase inhibitor valproic acid sensitizes human and canine osteosarcoma to doxorubicin. Cancer Chemother Pharmacol. 2011;67:83–92.

    CAS  PubMed  Google Scholar 

  42. Woronicz JD, Gao X, Cao Z, Rothe M, Goeddel DV. IkappaB kinase-beta: NF-kappaB activation and complex formation with IkappaB kinase-alpha and NIK. Science. 1997;278:866–9.

    CAS  PubMed  Google Scholar 

  43. Xiang M, Birkbak NJ, Vafaizadeh V, Walker SR, Yeh JE, Liu S, et al. STAT3 induction of miR-146b forms a feedback loop to inhibit the NF-kappaB to IL-6 signaling axis and STAT3-driven cancer phenotypes. Sci Signal. 2014;7:ra11.

    PubMed  PubMed Central  Google Scholar 

  44. Xiao H, Chen L, Luo G, Son H, Prectoni JH, Zheng W. Effect of the cytokine levels in serum on osteosarcoma. Tumour Biol. 2014;35:1023–8.

    CAS  PubMed  Google Scholar 

  45. Yang Q, Zage P, Kagan D, Tian Y, Seshadri R, Salwen HR, et al. Association of epigenetic inactivation of RASSF1A with poor outcome in human neuroblastoma. Clin Cancer Res. 2004;10:8493–500.

    CAS  PubMed  Google Scholar 

  46. Zhang L, Wang G, Wang L, Song C, Leng Y, Wang X, et al. VPA inhibits breast cancer cell migration by specifically targeting HDAC2 and down-regulating Survivin. Mol Cell Biochem. 2012;361:39–45.

    CAS  PubMed  Google Scholar 

  47. Zhang Z, Wang Y, Chen J, Tan Q, Xie C, Li C, et al. Silencing of histone deacetylase 2 suppresses malignancy for proliferation, migration, and invasion of glioblastoma cells and enhances temozolomide sensitivity. Cancer Chemother Pharmacol. 2016;78:1289–96.

    CAS  PubMed  Google Scholar 

Download references

Funding

This research was supported by the National Natural Science Foundation of China (Grant No. 81301546).

Author information

Affiliations

Authors

Corresponding authors

Correspondence to Wen Wu or Ming Liu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Li, J., Yan, X., Tang, J. et al. HDAC2-mediated upregulation of IL-6 triggers the migration of osteosarcoma cells. Cell Biol Toxicol 35, 423–433 (2019). https://doi.org/10.1007/s10565-019-09459-7

Download citation

Keywords

  • HDAC2
  • IL-6
  • Osteosarcoma
  • p65
  • IKK-β