Skip to main content

Advertisement

SpringerLink
Log in

Modulation of Regorafenib effects on HCC cell lines by epidermal growth factor

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

Blood platelet numbers are correlated to growth and aggressiveness of several tumor types, including hepatocellular carcinoma (HCC). We previously found that platelet lysates (hPLs) also stimulated growth and migration, and antagonized the growth-inhibitory and apoptotic effects of both Sorafenib and Regorafenib, two multikinase inhibitors, on three HCC cell lines. In this study, in vitro function of human epidermal growth factor (EGF) with and without Sorafenib or Regorafenib was investigated.

Methods

An ELISA kit was used to evaluate the EGF concentrations in hPLs. In vitro function of EGF was assessed with proliferation MTT test. Apoptosis assay, scratch assays, and Transwell assays were performed for apoptosis, invasion, and migration, respectively. MAPK Activation Kit was used to explore MAPK phosphorylation.

Results

EGF antagonized the growth inhibition of Regorafenib on three HCC cell lines. Regorafenib-mediated growth inhibition was blocked by 70 % when the cells were pre-treated with EGF. EGF also blocked Regorafenib-induced apoptosis, as well as Regorafenib-induced decreases in cell migration and invasion. The EGF effects were in turn antagonized by concomitant addition to the cultures of EGF receptor antagonist Erlotinib, showing that the EGF receptor was involved in the mechanisms of EGF-mediated blocking of Regorafenib effects. Erlotinib also partially blocked the effects of hPLs in antagonizing Regorafenib-mediated growth inhibition, showing that EGF was an important component of hPL actions.

Conclusions

All these results show that EGF antagonized Regorafenib-mediated growth and migration inhibition and apoptosis induction in HCC cells and reinforce the idea that microenvironment can influence cancer drug actions.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Levin J, Conley CL (1964) Thrombocytosis associated with malignant disease. Arch Intern Med 114:497–500

    Article  CAS  PubMed  Google Scholar 

  2. Stone RL, Nick AM, McNeish IA et al (2012) Paraneoplastic thrombocytosis in ovarian cancer. N Engl J Med 366:610–618

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Labelle M, Begum S, Hynes RO (2011) Direct signaling between platelets and cancer cells induces an epithelial-mesenchymal-like transition and promotes metastasis. Cancer Cell 20:576–590

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Holmes CE, Levis JE, Ornstein DL (2009) Activated platelets enhance ovarian cancer cell invasion in acellular model of metastasis. Clin Exp Metastasis 26:653–661

    Article  CAS  PubMed  Google Scholar 

  5. Steller EJ, Raats DA, Koster J et al (2013) PDGFRB promotes liver metastasis Formation of mesenchymal-like colorectal tumor cells. Neoplasia 15:204–207

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Hwang SJ, Luo JC, Li CP et al (2004) Thrombocytosis: a paraneoplastic syndrome in patients with hepatocellular carcinoma. World J Gastroenterol 10:2472–2477

    PubMed  Google Scholar 

  7. Carr BI, Guerra V (2013) Thrombocytosis and hepatocellular carcinoma. Dig Dis Sci 58:1790–1796

    Article  PubMed Central  PubMed  Google Scholar 

  8. Carr BI, Guerra V, Pancoska P (2012) Thrombocytopenia in relation to tumor size in patients with hepatocellular carcinoma. Oncology 83:339–345

    Article  CAS  PubMed  Google Scholar 

  9. Carr BI, Guerra V (2014) Features of massive hepatocellular carcinomas. Eur J Gastroenterol Hepatol 26:101–108

    Article  CAS  PubMed  Google Scholar 

  10. Carr BI, Cavallini A, D’Alessandro R et al (2014) Platelet extracts induce growth, migration and invasion in human hepatocellular carcinoma in vitro. BMC Cancer 14:351

    Article  PubMed Central  PubMed  Google Scholar 

  11. D’Alessandro R, Refolo MG, Lippolis C et al (2014) Antagonism of Sorafenib and Regorafenib actions by platelet factors in hepatocellular carcinoma cell lines. BMC Cancer 14:351

    Article  PubMed Central  PubMed  Google Scholar 

  12. Refolo MG, D’Alessandro R, Lippolis C et al (2014) Modulation of doxorubicin mediated growth inhibition of hepatocellular carcinoma cells by platelet lysates. Anticancer Agents Med Chem 14:1154–1160

    Article  CAS  PubMed  Google Scholar 

  13. Buergy D, Wenz F, Groden C, Brockmann MA (2012) Tumor-platelet interaction in solid tumors. Int J Cancer 130:2747–2760

    Article  CAS  PubMed  Google Scholar 

  14. Song S, Wientjes MG, Gan Y, Au JL (2000) Fibroblast growth factors: an epigenetic mechanism of broad spectrum resistance to anticancer drugs. Proc Natl Acad Sci USA 97:8658–8663

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Shen K, Cui D, Sun L, Lu Y, Han M (2012) Inhibition of IGF-IR increases chemosensitivity in human colorectal cancer cells through MRP-2 promoter suppression. J Cell Biochem 113:2086–2089

    Article  CAS  PubMed  Google Scholar 

  16. Jalving M, Gietema JA, Lefrandt JD, de Jong S, Reyners AK, Gans RO, de Vries EG (2010) Metformin: taking away the candy for cancer? Eur J Cancer 46(13):2369–2380

    Article  CAS  PubMed  Google Scholar 

  17. Atzori F, Traina TA, Ionta MT, Massidda B (2009) Targeting insulin-like growth factor type 1 receptor in cancer therapy. Target Oncol 4:255–266

    Article  PubMed  Google Scholar 

  18. Kina S, Phonaphonh T, Liang F et al (2013) PDGFα receptor is a mediator for cisplatin-induced Met expression. Eur J Pharmacol 699:227–232

    Article  CAS  PubMed  Google Scholar 

  19. Zhou T, Duan J, Wang Y et al (2012) Fluoxetine synergys with anticancer drugs to overcome multidrug resistance in breast cancer cells. Tumour Biol 33:1299–1306

    Article  CAS  PubMed  Google Scholar 

  20. Peer D, Dekel Y, Melikhov D, Margalit R (2004) Fluoxetine inhibits multidrug resistance extrusion pumps and enhances responses to chemotherapy in syngeneic and in human xenograft mouse tumor models. Cancer Res 64:7562–7569

    Article  CAS  PubMed  Google Scholar 

  21. Fuchs BC, Hoshida Y, Fujii T et al (2014) Epidermal growth factor receptor inhibition attenuates liver fibrosis and development of hepatocellular carcinoma. Hepatology 59:1577–1590

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Liang CC, Park AY, Guan JL (2007) In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2:329–333

    Article  CAS  PubMed  Google Scholar 

  23. Carr BI, D’Alessandro R, Refolo MG et al (2013) Effects of low concentrations of Regorafenib and Sorafenib on human HCC cell AFP, migration, invasion, and growth in vitro. J Cell Physiol 228:1344–1350

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Fan FT, Shen CS, Tao L et al (2014) PKM2 regulates hepatocellular carcinoma cell epithelial-mesenchymal transition and migration upon EGFR activation. Asian Pac J Cancer Prev 15:1961–1970

    Article  PubMed  Google Scholar 

  25. Hu Z, Du J, Yang L et al (2012) GEP100/Arf6 is required for epidermal growth factor-induced ERK/Rac1 signaling and cell migration in human hepatoma HepG2 cells. PLoS One 7:e38777

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Schallmoser K, Bartmann C, Rohde E et al (2007) Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells. Transfusion 47:1436–1446

    Article  CAS  PubMed  Google Scholar 

  27. Alexia C, Fallot G, Lasfer M, Schweizer-Groyer G, Groyer A (2004) An evaluation of the role of insulin-like growth factors (IGF) and of type-I IGF receptor signaling in hepatocarcinogenesis and in the resistance of HCC cells against drug-induced apoptosis. Biochem Pharmacol 68:1003–1015

    Article  CAS  PubMed  Google Scholar 

  28. Liang C, Chen W, Zhi X (2013) Serotonin promotes the proliferation of serum-deprived hepatocellular carcinoma cells via upregulation of FOXO3a. Mol Cancer 9:12–14

    Google Scholar 

  29. French DM, Lin BC, Wang M et al (2012) Targeting FGFR4 inhibits hepatocellular carcinoma in preclinical mouse models. PLoS One 7:e36713

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  30. Radziwon-Balicka A, Medina C, O’Driscoll L et al (2012) Platelets increase survival of adenocarcinoma cells challenged with anticancer drugs: mechanisms and implications for chemoresistance. Br J Pharmacol 167:787–804

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  31. Gilbert LA, Hemann MT (2011) Chemotherapeutic resistance: surviving stressful situations. Cancer Res 71:5062–5066

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  32. Tian X, Hao K, Qin C, Xie K, Xie X, Yang Y (2013) Insulin-like growth factor 1 receptor promotes the growth and chemoresistance of pancreatic cancer. Dig Dis Sci 58:2705–2712

    Article  CAS  PubMed  Google Scholar 

  33. Huang P, Xu X, Wang L, Zhu B, Wang X, Xia J (2014) The role of EGF-EGFR signalling pathway in hepatocellular carcinoma inflammatory microenvironment. J Cell Mol Med 18:218–230

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Carr BI, Cavallini A, Lippolis C et al (2013) Fluoro-Sorafenib (Regorafenib) effects on hepatoma cells: growth inhibition, quiescence, and recovery. J Cell Physiol 228:292–297

    Article  CAS  PubMed  Google Scholar 

  35. D’Alessandro R, Refolo MG, Lippolis C et al (2013) Reversibility of regorafenib effects in hepatocellular carcinoma cells. Cancer Chemother Pharmacol 72:869–877

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Italian Ministry of Public Health.

Conflict of interest

None.

Author information

Author notes

    Authors and Affiliations

    1. Laboratory of Cellular and Molecular Biology, Department Clinical Pathology, National Institute for Digestive Diseases, IRCCS “Saverio de Bellis”, Via Turi 27, 70013, Castellana Grotte, BA, Italy

      Rosalba D’Alessandro, Maria Grazia Refolo, Catia Lippolis, Nicola Carella, Caterina Messa, Aldo Cavallini & Brian Irving Carr

    2. Department of Gastroenterology and Hepatology, Sourasky Medical Center, Tel Aviv, Israel

      Rosalba D’Alessandro, Maria Grazia Refolo, Catia Lippolis, Nicola Carella, Caterina Messa, Aldo Cavallini & Brian Irving Carr

    Authors
    1. Rosalba D’Alessandro
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Maria Grazia Refolo
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Catia Lippolis
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Nicola Carella
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Caterina Messa
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Aldo Cavallini
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Brian Irving Carr
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Brian Irving Carr.

    Additional information

    Rosalba D’Alessandro and Maria Grazia Refolo have equally contributed.

    Professor Brian Irving Carr is on sabbatical leave.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    D’Alessandro, R., Refolo, M.G., Lippolis, C. et al. Modulation of Regorafenib effects on HCC cell lines by epidermal growth factor. Cancer Chemother Pharmacol 75, 1237–1245 (2015). https://doi.org/10.1007/s00280-015-2751-6

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00280-015-2751-6

    Keywords

    Search

    Navigation