Immunologic Research

, Volume 67, Issue 6, pp 505–516 | Cite as

Blockade of TLR4 using TAK-242 (resatorvid) enhances anti-cancer effects of chemotherapeutic agents: a novel synergistic approach for breast and ovarian cancers

  • Bahareh Kashani
  • Zahra Zandi
  • Mohammad Reza Karimzadeh
  • Davood Bashash
  • Ali Nasrollahzadeh
  • Seyed H. GhaffariEmail author
Original Article


It is believed that pathways of the immune system are responsible for eradicating cancer cells; however, their over-activation and also their ectopic expression in tumor cells and microenvironment are major contributors to tumor growth and chemoresistance. Toll-like receptor 4 (TLR4) pathway is an innate immune-related pathway which is usually overexpressed in tumor cells that leads to excessive pro-inflammatory cytokines and eventually results in tumor survival, drug resistance, and metastasis. In this study, we investigated whether TLR4 expression is affected upon the treatment of breast and ovarian cancer cells with common chemotherapeutics (paclitaxel, cisplatin, doxorubicin, and arsenic trioxide) and if TLR4 inhibition using its specific inhibitor TAK-242 could enhance cancer cells’ response to the drugs. Both breast (MCF7) and ovarian (2008C13) cancer cells experienced an elevated expression of TLR4 after treatment with the drugs. The expression of this receptor was also upregulated in cisplatin-resistant 2008C13 cells; however, it was significantly higher upon short-term treatment with cisplatin. More importantly, the combination treatment of the drugs with TAK-242 intensified the chemosensitivity of six different breast and ovarian cancer cells to chemotherapeutic drugs. It was also identified that co-treatment of paclitaxel and TAK-242 not only led to enhanced G2/M arrest and apoptosis but also satisfactorily decreased the expression of TLR4 and different interleukins in these cells. Taken together, the results of the present study emphasize that chemotherapy may lead to chemoresistance through inducing TLR4 expression, and therefore inhibiting this receptor using TAK-242 could be a promising approach to improve the outcome of chemotherapy in foreseeable future.


Toll-like receptor 4 (TLR4) TAK-242 Chemotherapy Combination therapy Breast and ovarian cancers 



The authors would like to acknowledge the help provided by the Medical University of Bam.

Funding information

Research reported in this publication was supported by a grant from Hematology, Oncology, and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Informed consent


Research involving human participants and/or animals

Not applicable

Supplementary material

12026_2019_9113_MOESM1_ESM.docx (687 kb)
ESM 1 (DOCX 687 KB)


  1. 1.
    Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394–424.Google Scholar
  2. 2.
    Doubeni CA, Doubeni AR, Myers AE, Doubeni AR. Diagnosis and Management of Ovarian Cancer. American family physician. 2016 Jun 1;93(11).Google Scholar
  3. 3.
    Yates LR, Knappskog S, Wedge D, et al. Genomic evolution of breast cancer metastasis and relapse. Cancer Cell. 2017;32(2):169–84. e7.PubMedPubMedCentralGoogle Scholar
  4. 4.
    Ye X, Brabletz T, Kang Y, et al. Upholding a role for EMT in breast cancer metastasis. Nature. 2017;547(7661):E1.PubMedPubMedCentralGoogle Scholar
  5. 5.
    Holmes D. Ovarian cancer: beyond resistance. Nature. 2015;527(7579):S217.PubMedGoogle Scholar
  6. 6.
    Dagogo-Jack I, Shaw AT. Tumour heterogeneity and resistance to cancer therapies. Nat Rev Clin Oncol. 2018;15(2):81–94.PubMedGoogle Scholar
  7. 7.
    Moscow JA, Cowan KH, Sikic BI. Drug resistance and its clinical circumvention. Holland‐Frei Cancer Medicine. 2016 Oct 17:1-7.Google Scholar
  8. 8.
    Lipinski KA, Barber LJ, Davies MN, Ashenden M, Sottoriva A, Gerlinger M. Cancer evolution and the limits of predictability in precision cancer medicine. Trends Cancer. 2016;2(1):49–63.PubMedPubMedCentralGoogle Scholar
  9. 9.
    Janssen LM, Ramsay EE, Logsdon CD, Overwijk WW. The immune system in cancer metastasis: friend or foe? J Immunother Cancer. 2017;5(1):79.PubMedPubMedCentralGoogle Scholar
  10. 10.
    Keibel A, Singh V, Sharma MC. Inflammation, microenvironment, and the immune system in cancer progression. Curr Pharm Des. 2009;15(17):1949–55.PubMedGoogle Scholar
  11. 11.
    Chung YH, Kim D. Enhanced TLR4 expression on colon cancer cells after chemotherapy promotes cell survival and epithelial–mesenchymal transition through phosphorylation of GSK3β. Anticancer Res. 2016;36(7):3383–94.PubMedGoogle Scholar
  12. 12.
    Lu Y-C, Yeh W-C, Ohashi PS. LPS/TLR4 signal transduction pathway. Cytokine. 2008;42(2):145–51.PubMedGoogle Scholar
  13. 13.
    Ara T, DeClerck YA. Interleukin-6 in bone metastasis and cancer progression. Eur J Cancer. 2010;46(7):1223–31.PubMedPubMedCentralGoogle Scholar
  14. 14.
    Mai CW, Kang YB, Pichika MR. Should a toll-like receptor 4 (TLR-4) agonist or antagonist be designed to treat cancer? TLR-4: its expression and effects in the ten most common cancers. Onco Targets Ther. 2013;6:1573.PubMedPubMedCentralGoogle Scholar
  15. 15.
    Szajnik M, Szczepanski MJ, Czystowska M, Elishaev E, Mandapathil M, Nowak-Markwitz E, et al. TLR4 signaling induced by lipopolysaccharide or paclitaxel regulates tumor survival and chemoresistance in ovarian cancer. Oncogene. 2009;28(49):4353–63.PubMedPubMedCentralGoogle Scholar
  16. 16.
    Rovere-Querini P, Castiglioni A. Adjuvant role for cell death during chemo- and radiotherapy of cancer? Expert Rev Clin Immunol. 2008;4(1):27–32.PubMedGoogle Scholar
  17. 17.
    Zhou L, Qi L, Jiang L, Zhou P, Ma J, Xu X, et al. Antitumor activity of gemcitabine can be potentiated in pancreatic cancer through modulation of TLR4/NF-κB signaling by 6-shogaol. AAPS J. 2014;16(2):246–57.PubMedPubMedCentralGoogle Scholar
  18. 18.
    Kawamoto T, Ii M, Kitazaki T, Iizawa Y, Kimura H. TAK-242 selectively suppresses toll-like receptor 4-signaling mediated by the intracellular domain. Eur J Pharmacol. 2008;584(1):40–8.PubMedGoogle Scholar
  19. 19.
    Matsunaga N, Tsuchimori N, Matsumoto T, Ii M. TAK-242 (resatorvid), a small-molecule inhibitor of toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules. Mol Pharmacol. 2011;79(1):34–41.PubMedGoogle Scholar
  20. 20.
    Sha T, Iizawa Y, Ii M. Combination of imipenem and TAK-242, a toll-like receptor 4 signal transduction inhibitor, improves survival in a murine model of polymicrobial sepsis. Shock. 2011;35(2):205–9.PubMedGoogle Scholar
  21. 21.
    Fenhammar J, Rundgren M, Forestier J, Kalman S, Eriksson S, Frithiof R. Toll-like receptor 4 inhibitor TAK-242 attenuates acute kidney injury in endotoxemic sheep. Anesthesiology. 2011;114(5):1130–7.PubMedGoogle Scholar
  22. 22.
    Okada T, Lei L, Nishikawa H, Nakano F, Nakatsuka Y, Suzuki H. TAK-242, toll-like receptor 4 antagonist, attenuates brain edema in subarachnoid hemorrhage mice. Subarachnoid Hemorrhage: Springer; 2020. p. 77–81.Google Scholar
  23. 23.
    Zandi Z, Kashani B, Bashash D, Poursani EM, Mousavi SA, Chahardoli B, Ghaffari SH. The anticancer effect of the TLR4 inhibition using TAK‐242 (resatorvid) either as a single agent or in combination with chemotherapy: A novel therapeutic potential for breast cancer. Journal of cellular biochemistry. 2019 Sep 18.Google Scholar
  24. 24.
    Kashani B, Zandi Z, Bashash D, Zaghal A, Momeny M, Poursani EM, Pourbagheri-Sigaroodi A, Mousavi SA, Ghaffari SH. Small molecule inhibitor of TLR4 inhibits ovarian cancer cell proliferation: new insight into the anticancer effect of TAK-242 (Resatorvid). Cancer Chemotherapy and Pharmacology. 2019 Nov 30:1-3.Google Scholar
  25. 25.
    Zandi Z, Kashani B, Poursani EM, Bashash D, Kabuli M, Momeny M, et al. TLR4 blockade using TAK-242 suppresses ovarian and breast cancer cells invasion through the inhibition of extracellular matrix degradation and epithelial-mesenchymal transition. Eur J Pharmacol. 2019;853:256–63. Scholar
  26. 26.
    Chou T-C. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010:0008–5472. CAN-09-1947.Google Scholar
  27. 27.
    Yuan X, Zhou Y, Wang W, et al. Activation of TLR4 signaling promotes gastric cancer progression by inducing mitochondrial ROS production. Cell Death Dis. 2013;4(9):e794.PubMedPubMedCentralGoogle Scholar
  28. 28.
    Apetoh L, Ghiringhelli F, Tesniere A, et al. The interaction between HMGB1 and TLR4 dictates the outcome of anticancer chemotherapy and radiotherapy. Immunol Rev. 2007;220(1):47–59.PubMedGoogle Scholar
  29. 29.
    Zeineddine D, Hammoud AA, Mortada M, Boeuf H. The Oct4 protein: more than a magic stemness marker. Am J Stem Cells. 2014;3(2):74.PubMedPubMedCentralGoogle Scholar
  30. 30.
    Chang T-S, Chen C-L, Wu Y-C, Liu JJ, Kuo YC, Lee KF, et al. Inflammation promotes expression of stemness-related properties in HBV-related hepatocellular carcinoma. PLoS One. 2016;11(2):e0149897.PubMedPubMedCentralGoogle Scholar
  31. 31.
    Hagemann T, Balkwill F, Lawrence T. Inflammation and cancer: a double-edged sword. Cancer Cell. 2007;12(4):300–1.PubMedPubMedCentralGoogle Scholar
  32. 32.
    Balkwill F. Cancer and the chemokine network. Nat Rev Cancer. 2004;4(7):540–50.PubMedPubMedCentralGoogle Scholar
  33. 33.
    He W, Liu Q, Wang L, Chen W, Li N, Cao X. TLR4 signaling promotes immune escape of human lung cancer cells by inducing immunosuppressive cytokines and apoptosis resistance. Mol Immunol. 2007;44(11):2850–9.PubMedGoogle Scholar
  34. 34.
    Brunialti MKC, Martins PS, de Carvalho HB, Machado FR, Barbosa LM, Salomao R. TLR2, TLR4, CD14, CD11B, and CD11C expressions on monocytes surface and cytokine production in patients with sepsis, severe sepsis, and septic shock. Shock. 2006;25(4):351–7.PubMedGoogle Scholar
  35. 35.
    Sabroe I, Parker L, Wilson A, Whyte MB, Dower S. Toll-like receptors: their role in allergy and non-allergic inflammatory disease. Clin Exp Allergy. 2002;32(7):984–9.PubMedGoogle Scholar
  36. 36.
    González-Reyes S, Marín L, González L, et al. Study of TLR3, TLR4 and TLR9 in breast carcinomas and their association with metastasis. BMC Cancer. 2010;10(1):665.PubMedPubMedCentralGoogle Scholar
  37. 37.
    Luo X-Z, He Q-Z, Wang K. Expression of toll-like receptor 4 in ovarian serous adenocarcinoma and correlation with clinical stage and pathological grade. Int J Clin Exp Med. 2015;8(8):14323–7.PubMedPubMedCentralGoogle Scholar
  38. 38.
    Szczepanski MJ, Czystowska M, Szajnik M, Harasymczuk M, Boyiadzis M, Kruk-Zagajewska A, et al. Triggering of toll-like receptor 4 expressed on human head and neck squamous cell carcinoma promotes tumor development and protects the tumor from immune attack. Cancer Res. 2009;69(7):3105–13.PubMedPubMedCentralGoogle Scholar
  39. 39.
    Sheyhidin I, Nabi G, Hasim A, Zhang RP, Ainiwaer J, Ma H, et al. Overexpression of TLR3, TLR4, TLR7 and TLR9 in esophageal squamous cell carcinoma. World J Gastroenterol: WJG. 2011;17(32):3745–51.PubMedGoogle Scholar
  40. 40.
    Manček-Keber M, Jerala R. Postulates for validating TLR4 agonists. Eur J Immunol. 2015;45(2):356–70.PubMedGoogle Scholar
  41. 41.
    Zimmer SM, Liu J, Clayton JL, Stephens DS, Snyder JP. Paclitaxel binding to human and murine MD-2. J Biol Chem. 2008;283(41):27916–26.PubMedPubMedCentralGoogle Scholar
  42. 42.
    Lv W, Chen N, Lin Y, et al. Macrophage migration inhibitory factor promotes breast cancer metastasis via activation of HMGB1/TLR4/NF kappa B axis. Cancer Lett. 2016;375(2):245–55.PubMedGoogle Scholar
  43. 43.
    Ren B, Luo S, Tian X, Jiang Z, Zou G, Xu F, et al. Curcumin inhibits liver cancer by inhibiting DAMP molecule HSP70 and TLR4 signaling. Oncol Rep. 2018;40(2):895–901.PubMedGoogle Scholar
  44. 44.
    Zhou S, Du R, Wang Z, Shen W, Gao R, Jiang S, Fang Y, Shi Y, Chang A, Liu L, Liu C. TLR 4 increases the stemness and is highly expressed in relapsed human hepatocellular carcinoma. Cancer medicine. 2019 May 1.Google Scholar
  45. 45.
    Rothchild, Greg A., George Sandusky, Constance J. Temm, and Costantine Albany. "Identifying a predictive biomarker for chemotherapy (taxane) in patients with metastatic castration resistance prostate cancer." (2016): 426-426.Google Scholar
  46. 46.
    Rajput S, Volk-Draper LD, Ran S. TLR4 is a novel determinant of the response to paclitaxel in breast cancer. Mol Cancer Ther. 2013;12(8):1676–87.PubMedPubMedCentralGoogle Scholar
  47. 47.
    Wang A, Su Q, Wu F, Zhang X, Liu P. Role of TLR4 for paclitaxel chemotherapy in human epithelial ovarian cancer cells. Eur J Clin Investig. 2009;39(2):157–64.Google Scholar
  48. 48.
    Volk-Draper L, Hall K, Griggs C, Rajput S, Kohio P, DeNardo D, et al. Paclitaxel therapy promotes breast cancer metastasis in a TLR4-dependent manner. Cancer Res. 2014;74(19):5421–34.PubMedPubMedCentralGoogle Scholar
  49. 49.
    Sun NK, Huang SL, Chang TC, Chao CCK. TLR4 and NFκB signaling is critical for taxol resistance in ovarian carcinoma cells. J Cell Physiol. 2018;233(3):2489–501.PubMedGoogle Scholar
  50. 50.
    Huang J-M, Zhang G-N, Shi Y, et al. Atractylenolide-I sensitizes human ovarian cancer cells to paclitaxel by blocking activation of TLR4/MyD88-dependent pathway. Sci Rep. 2014;4:3840.PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, School of MedicineTehran University of Medical SciencesTehranIran
  2. 2.Department of Medical Genetics, School of MedicineTehran University of Medical SciencesTehranIran
  3. 3.Department of Medical Genetics, School of MedicineBam University of Medical SciencesBamIran
  4. 4.Department of Hematology and Blood Banking, School of Allied Medical SciencesShahid Beheshti University of Medical SciencesTehranIran

Personalised recommendations