Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Diacerein-mediated inhibition of IL-6/IL-6R signaling induces apoptotic effects on breast cancer

Oncogene volume 35, pages 3965–3975 (2016)Cite this article

Subjects

Abstract

Interleukin-6 (IL-6) signaling network has been implicated in oncogenic transformations making it attractive target for the discovery of novel cancer therapeutics. In this study, potent antiproliferative and apoptotic effect of diacerein were observed against breast cancer. In vitro apoptosis was induced by this drug in breast cancer cells as verified by increased sub-G1 population, LIVE/DEAD assay, cell cytotoxicity and presence of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells, as well as downregulation of antiapoptotic proteins Bcl-2 and Bcl-xL and upregulation of apoptotic protein Bax. In addition, apoptosis induction was found to be caspase dependent. Further molecular investigations indicated that diacerein instigated apoptosis was associated with inhibition of IL-6/IL-6R autocrine signaling axis. Suppression of STAT3, MAPK and Akt pathways were also observed as a consequence of diacerein-mediated upstream inhibition of IL-6/IL-6R. Fluorescence study and western blot analysis revealed cytosolic accumulation of STAT3 in diacerein-treated cells. The docking study showed diacerein/IL-6R interaction that was further validated by competitive binding assay and isothermal titration calorimetry. Most interestingly, it was found that diacerein considerably suppressed tumor growth in MDA-MB-231 xenograft model. The in vivo antitumor effect was correlated with decreased proliferation (Ki-67), increased apoptosis (TUNEL) and inhibition of IL-6/IL-6R-mediated STAT3, MAPK and Akt pathway in tumor remnants. Taken together, diacerein offered a novel blueprint for cancer therapy by hampering IL-6/IL-6R/STAT3/MAPK/Akt network.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Ghofrani HA, Osterloh IH, Grimminger F . Sildenafil: from angina to erectile dysfunction to pulmonary hypertension and beyond. Nat Rev Drug Discov 2006; 5: 689–702.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Palumbo A, Facon T, Sonneveld P, Blade J, Offidani M, Gay F et al. Thalidomide for treatment of multiple myeloma: 10 years later. Blood 2008; 111: 3968–3977.

    Article  CAS  PubMed  Google Scholar 

  3. Pooja S, Chaudhary P, Nayak LV, Rajender S, Saini KS, Deol D et al. Polymorphic variations in IL-1beta, IL-6 and IL-10 genes, their circulating serum levels and breast cancer risk in Indian women. Cytokine 2012; 60: 122–128.

    Article  CAS  PubMed  Google Scholar 

  4. Sachdev JC, Jahanzeb M . Blockade of the HER family of receptors in the treatment of HER2-positive metastatic breast cancer. Clin Breast Cancer 2012; 12: 19–29.

    Article  CAS  PubMed  Google Scholar 

  5. Culig Z . Cytokine disbalance in common human cancers. Biochim Biophys Acta 2011; 1813: 308–314.

    Article  CAS  PubMed  Google Scholar 

  6. Witsch E, Sela M, Yarden Y . Roles for growth factors in cancer progression. Physiology 2010; 25: 85–101.

    Article  CAS  PubMed  Google Scholar 

  7. Colotta F, Allavena P, Sica A, Garlanda C, Mantovani A . Cancer-related inflammation, the seventh hallmark of cancer: links to genetic instability. Carcinogenesis 2009; 30: 1073–1081.

    Article  CAS  PubMed  Google Scholar 

  8. Silver JS, Hunter CA . gp130 at the nexus of inflammation, autoimmunity, and cancer. J Leukocyte Biol 2010; 88: 1145–1156.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Gibson RM, Schiemann WP, Prichard LB, Reno JM, Ericsson LH, Nathanson NM . Phosphorylation of human gp130 at Ser-782 adjacent to the di-leucine internalization motif - Effects on expression and signaling. J Biol Chem 2000; 275: 22574–22582.

    Article  CAS  PubMed  Google Scholar 

  10. Jones SA, Scheller J, Rose-John S . Therapeutic strategies for the clinical blockade of IL-6/gp130 signaling. J Clin Invest 2011; 121: 3375–3383.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hideshima T, Nakamura N, Chauhan D, Anderson KC . Biologic sequelae of interleukin-6 induced PI3-K/Akt signaling in multiple myeloma. Blood 2001; 98: 639a.

    Google Scholar 

  12. Cavarretta IT, Neuwirt H, Untergasser G, Moser PL, Zaki MH, Steiner H et al. The antiapoptotic effect of IL-6 autocrine loop in a cellular model of advanced prostate cancer is mediated by Mcl-1. Oncogene 2007; 26: 2822–2832.

    Article  CAS  PubMed  Google Scholar 

  13. Lu ZM, Xu SC . ERK1/2 MAP kinases in cell survival and apoptosis. IUBMB Life 2006; 58: 621–631.

    Article  CAS  PubMed  Google Scholar 

  14. Rawlings JS, Rosler KM, Harrison DA . The JAK/STAT signaling pathway. J Cel Sci 2004; 117: 1281–1283.

    Article  CAS  Google Scholar 

  15. Dolled-Filhart M, Camp RL, Kowalski DP, Smith BL, Rimm DL . Tissue microarray analysis of signal transducers and activators of transcription 3 (Stat3) and Phospho-Stat3 (Tyr705) in node-negative breast cancer shows nuclear localization is associated with a better prognosis. Clin Cancer Res 2003; 9: 594–600.

    CAS  PubMed  Google Scholar 

  16. Puthier D, Derenne S, Barille S, Moreau P, Harousseau JL, Bataille R et al. Mcl-1 and Bcl-xL are co-regulated by IL-6 in human myeloma cells. Br J Haematol 1999; 107: 392–395.

    Article  CAS  PubMed  Google Scholar 

  17. Bollrath J, Phesse TJ, von Burstin VA, Putoczki T, Bennecke M, Bateman T et al. gp130-mediated Stat3 activation in enterocytes regulates cell survival and cell-cycle progression during colitis-associated tumorigenesis. Cancer Cell 2009; 15: 91–102.

    Article  CAS  PubMed  Google Scholar 

  18. Guo YQ, Xu F, Lu TH, Duan ZF, Zhang Z . Interleukin-6 signaling pathway in targeted therapy for cancer. Cancer Treat Rev 2012; 38: 904–910.

    Article  CAS  PubMed  Google Scholar 

  19. Berishaj M, Gao SP, Ahmed S, Leslie K, Al-Ahmadie H, Gerald WL et al. Stat3 is tyrosine-phosphorylated through the interleukin-6/glycoprotein 130/Janus kinase pathway in breast cancer. Br Cancer Res 2007; 9: R32.

    Article  Google Scholar 

  20. Shariat SF, Chromecki TF, Hoefer J, Barbieri CE, Scherr DS, Karakiewicz PI et al. Soluble gp130 regulates prostate cancer invasion and progression in an interleukin-6 dependent and independent manner. J Urol 2011; 186: 2107–2114.

    Article  CAS  PubMed  Google Scholar 

  21. Rokavec M, Oner MG, Li H, Jackstadt R, Jiang L, Lodygin D et al. IL-6R/STAT3/miR-34a feedback loop promotes EMT-mediated colorectal cancer invasion and metastasis. J Clin Invest 2014; 124: 1853–1867.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Conze D, Weiss L, Regen PS, Bhushan A, Weaver D, Johnson P et al. Autocrine production of interleukin 6 causes multidrug resistance in breast cancer cells. Cancer Res 2001; 61: 8851–8858.

    CAS  PubMed  Google Scholar 

  23. Rose-John S . GP130 stimulation and the maintenance of stem cells. Trends Biotechnol 2002; 20: 417–419.

    Article  CAS  PubMed  Google Scholar 

  24. Calisto KL, Camacho AC, Mittestainer FC, Carvalho BM, Guadagnini D, Carvalheira JB et al. Diacerhein attenuates the inflammatory response and improves survival in a model of severe sepsis. Crit Care 2012; 16: R158.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Gadotti VM, Martins DF, Pinto HF, Oliveira G, Kaster MP, Quintao NLM et al. Diacerein decreases visceral pain through inhibition of glutamatergic neurotransmission and cytokine signaling in mice. Pharmacol Biochem Behav 2012; 102: 549–554.

    Article  CAS  PubMed  Google Scholar 

  26. Varghese JN, Moritz RL, Lou MZ, Van Donkelaar A, Ji H, Ivancic N et al. Structure of the extracellular domains of the human interleukin-6 receptor alpha -chain. Proc Natl Acad Sci USA 2002; 99: 15959–15964.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Michaud-Levesque J, Bousquet-Gagnon N, Beliveau R . Quercetin abrogates IL-6/STAT3 signaling and inhibits glioblastoma cell line growth and migration. Exp Cell Res 2012; 318: 925–935.

    Article  CAS  PubMed  Google Scholar 

  28. Ataie-Kachoie P, Pourgholami MH, Morris DL . Inhibition of the IL-6 signaling pathway: a strategy to combat chronic inflammatory diseases and cancer. Cytokine Growth Factor Rev 2013; 24: 163–173.

    Article  CAS  PubMed  Google Scholar 

  29. Liu Y, Li PK, Li CL, Lin JY . Inhibition of STAT3 signaling blocks the anti-apoptotic activity of IL-6 in human liver cancer cells. J Biol Chem 2010; 285: 27429–27439.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Kamran MZ, Patil P, Gude RP . Role of STAT3 in cancer metastasis and translational advances. Biomed Res Int 2013; 2013: 421821.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Yosifov DY, Reufsteck C, Konstantinov SM, Berger MR . Interleukin-6, osteopontin and Raf/MEK/ERK signaling modulate the sensitivity of human myeloma cells to alkylphosphocholines. Leukemia Res 2012; 36: 764–772.

    Article  CAS  Google Scholar 

  32. Voorhees PM, Chen Q, Kuhn DJ, Small GW, Hunsucker SA, Strader JS et al. Inhibition of interleukin-6 signaling with CNTO 328 enhances the activity of bortezomib in preclinical models of multiple myeloma. Clin Cancer Res 2007; 13: 6469–6478.

    Article  CAS  PubMed  Google Scholar 

  33. Liu Y, Liu AG, Li HM, Li CL, Lin JY . Celecoxib inhibits interleukin-6/interleukin-6 receptor-induced JAK2/STAT3 phosphorylation in human hepatocellular carcinoma cells. Cancer Prev Res 2011; 4: 1296–1305.

    Article  CAS  Google Scholar 

  34. Brinkmann K, Kashkar H . Targeting the mitochondrial apoptotic pathway: a preferred approach in hematologic malignancies? Cell Death Dis 2014; 5: e1098.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Johnson C, Han Y, Hughart N, McCarra J, Alpini G, Meng F . Interleukin-6 and its receptor, key players in hepatobiliary inflammation and cancer. Transl Gastrointest Cancer 2012; 1: 58–70.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Ara T, DeClerck YA . Interleukin-6 in bone metastasis and cancer progression. Eur J Cancer 2010; 46: 1223–1231.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Liu YX, Law BK, Luesch H, Apratoxin A . Reversibly inhibits the secretory pathway by preventing cotranslational translocation. Mol Pharmacol 2009; 76: 91–104.

    Article  CAS  PubMed  Google Scholar 

  38. Chakravarti N, Myers JN, Aggarwal BB . Targeting constitutive and interleukin-6-inducible signal transducers and activators of transcription 3 pathway in head and neck squamous cell carcinoma cells by curcumin (diferuloylmethane). Int J Cancer 2006; 119: 1268–1275.

    Article  CAS  PubMed  Google Scholar 

  39. Bharti AC, Donato N, Aggarwal BB . Curcumin (diferuloylmethane) inhibits constitutive and IL-6-inducible STAT3 phosphorylation in human multiple myeloma cells. J Immunol 2003; 171: 3863–3871.

    Article  CAS  PubMed  Google Scholar 

  40. Plotnikov A, Zehorai E, Procaccia S, Seger R . The MAPK cascades: signaling components, nuclear roles and mechanisms of nuclear translocation. Biochim Biophys Acta 2011; 1813: 1619–1633.

    Article  CAS  PubMed  Google Scholar 

  41. Andersen JL, Schroder TJ, Christensen S, Strandbygard D, Pallesen LT, Garcia-Alai MM et al. Identification of the first small-molecule ligand of the neuronal receptor sortilin and structure determination of the receptor-ligand complex. Acta Crystallogr D Biol Crystallogr 2014; 70: 451–460.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Mahata D, Mandal SM, Bharti R, Gupta VK, Mandal M, Nag A et al. Self-assembled cardanol azo derivatives as antifungal agent with chitin-binding ability. Int J Biol Macromol 2014;69: 5–11.

    Article  CAS  PubMed  Google Scholar 

  43. Sarkar S, Mazumdar A, Dash R, Sarkar D, Fisher PB, Mandal M . ZD6474 enhances paclitaxel antiproliferative and apoptotic effects in breast carcinoma cells. J Cel Physiol 2011; 226: 375–384.

    Article  CAS  Google Scholar 

  44. Das S, Dey KK, Bharti R, MaitiChoudhury S, Maiti S, Mandal M . PKI 166 induced redox signalling and apoptosis through activation of p53, MAP kinase and caspase pathway in epidermoid carcinoma. J Exp Ther Oncol 2012; 10: 139–153.

    CAS  PubMed  Google Scholar 

  45. Rajput S, BNP Kumar, Dey KK, Pal oI, Parekh A, Mandal M . Molecular targeting of Akt by thymoquinone promotes G(1) arrest through translation inhibition of cyclin D1 and induces apoptosis in breast cancer cells. Life Sci 2013; 93: 783–790.

    Article  CAS  PubMed  Google Scholar 

  46. Venkatesan P, Puvvada N, Dash R, Kumar BNP, Sarkar D, Azab B et al. The potential of celecoxib-loaded hydroxyapatite-chitosan nanocomposite for the treatment of colon cancer. Biomaterials 2011; 32: 3794–3806.

    Article  CAS  PubMed  Google Scholar 

  47. Rajput S, Kumar BNP, Sarkar S, Das S, Azab B, Santhekadur PK et al. Targeted apoptotic effects of thymoquinone and tamoxifen on XIAP mediated Akt regulation in breast cancer. PLoS One 2013; 8: e61342.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Venkatesan SK, Shukla AK, Dubey VK . Molecular docking studies of selected tricyclic and quinone derivatives on trypanothione reductase of leishmania infantum. J Comput Chem 2010; 31: 2463–2475.

    CAS  PubMed  Google Scholar 

  49. Su JL, Lai KP, Chen CA, Yang CY, Chen PS, Chang CC et al. A novel peptide specifically binding to interleukin-6 receptor (gp80) inhibits angiogenesis and tumor growth. Cancer Res 2005; 65: 4827–4835.

    Article  CAS  PubMed  Google Scholar 

  50. Mandal M, Adam L, Mendelsohn J, Kumar R . Nuclear targeting of Bax during apoptosis in human colorectal cancer cells. Oncogene 1998; 17: 999–1007.

    Article  CAS  PubMed  Google Scholar 

  51. Sethi G, Chatterjee S, Rajendran P, Li F, Shanmugam MK, Wong KF et al. Inhibition of STAT3 dimerization and acetylation by garcinol suppresses the growth of human hepatocellular carcinoma in vitro and in vivo. Mol Cancer 2014; 13: 66.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Dey G, Bharti R, Dhanarajan G, Das S, Dey KK, Kumar BN et al. Marine lipopeptide Iturin A inhibits Akt mediated GSK3beta and FoxO3a signaling and triggers apoptosis in breast cancer. Sci Rep 2015; 5: 10316.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank MHRD, CSIR and MoES, India (Grant No: MoES/16/48/09/RDEAS; Date: 15-07-2010).

Author information

Authors and Affiliations

  1. School of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur, India

    R Bharti, G Dey, S Rajput & M Mandal

  2. Division of Medicinal and Pharmaceutical Chemistry, Department of Pharmaceutical Technology, Drug Theoretics and Cheminformatics Laboratory, Jadavpur University, Kolkata, India

    P K Ojha

  3. IJN-UTM Cardiovascular Engineering Centre, Faculty of Biosciences and Medical Engineering, Universiti, Teknologi, Malaysia

    S K Jaganathan

  4. Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, India

    R Sen

Authors
  1. R Bharti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G Dey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P K Ojha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Rajput
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S K Jaganathan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. R Sen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M Mandal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M Mandal.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bharti, R., Dey, G., Ojha, P. et al. Diacerein-mediated inhibition of IL-6/IL-6R signaling induces apoptotic effects on breast cancer. Oncogene 35, 3965–3975 (2016). https://doi.org/10.1038/onc.2015.466

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2015.466

This article is cited by

Search

Advanced search

Quick links