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RANKL-induced c-Src activation contributes to conventional anti-cancer drug resistance and dasatinib overcomes this resistance in RANK-expressing multiple myeloma cells

  • Keiji Mashimo
  • Masanobu Tsubaki
  • Tomoya Takeda
  • Ryota Asano
  • Minami Jinushi
  • Motohiro Imano
  • Takao Satou
  • Katsuhiko Sakaguchi
  • Shozo Nishida
Original Article

Abstract

The survival and growth of multiple myeloma (MM) cells are facilitated by cell–cell interactions with bone marrow stromal cells and the bone marrow microenvironment. These interactions induce de novo drug resistance known as cell adhesion-mediated drug resistance. Our previous results recently revealed that the receptor activator of NF-κB (RANK) ligand (RANKL), which is expressed by bone marrow stromal cells, contributes to anti-cancer drug resistance through the activation of various signaling molecules and suppression of Bim expression in RANK-expressing MM cells. However, the detailed mechanisms underlying RANKL-induced drug resistance remain uncharacterized. In the present study, we investigated the mechanism of RANKL-induced drug resistance in RANK-expressing MM cell lines. We found treatment of MM cells with RANKL-induced c-Src phosphorylation and activation of the downstream signaling molecules Akt, mTOR, STAT3, JNK, and NF-κB. In addition, treatment with dasatinib, a c-Src inhibitor, overcame RANKL- and bone marrow stromal cell-induced drug resistance to adriamycin, vincristine, dexamethasone, and melphalan by suppressing c-Src, Akt, mTOR, STAT3, JNK, and NF-κB activation and enhancing expression of Bim. Overall, RANKL- and bone marrow stromal cell-induced drug resistance correlated with the activation of c-Src signaling pathways, which caused a decrease in Bim expression. Dasatinib treatment of RANK-expressing MM cells re-sensitized them to anti-cancer drugs. Therefore, inhibition of c-Src may be a new therapeutic approach for overcoming RANKL-induced drug resistance in patients with MM.

Keywords

Multiple myeloma RANK RANKL Src Drug resistance 

Notes

Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research (C) (Grant Number 15K08116), Grant-in-Aid for Young Scientists (B) (Grant Number 16K18965) from the Japan Society for the Promotion of Science (JSPS) and by Ministry of Education, Culture, Sports, Science, and Technology (MEXT)-Supported Program for the Strategic Research Foundation at Private Universities, 2014-2018 (Grant number S1411037).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10238_2018_531_MOESM1_ESM.pdf (31 kb)
Supplementary material 1 (PDF 31 kb)

References

  1. 1.
    Ludwig H, Milosavljevic D, Zojer N, et al. Immunoglobulin heavy/light chain ratios improve paraprotein detection and monitoring, identify residual disease and correlate with survival in multiple myeloma patients. Leukemia. 2013;27(1):213–9.CrossRefGoogle Scholar
  2. 2.
    Tsubaki M, Satou T, Itoh T, et al. Overexpression of MDR1 and survivin, and decreased Bim expression mediate multidrug-resistance in multiple myeloma cells. Leuk Res. 2012;36(10):1315–22.CrossRefGoogle Scholar
  3. 3.
    Kumar SK, Rajkumar SV, Dispenzieri A, et al. Improved survival in multiple myeloma and the impact of novel therapies. Blood. 2008;111(5):2516–20.CrossRefGoogle Scholar
  4. 4.
    Mitsiades CS, Hayden PJ, Anderson KC, et al. From the bench to the bedside: emerging new treatments in multiple myeloma. Best Pract Res Clin Haematol. 2007;20(4):797–816.CrossRefGoogle Scholar
  5. 5.
    Meads MB, Hazlehurst LA, Dalton WS. The bone marrow microenvironment as a tumor sanctuary and contributor to drug resistance. Clin Cancer Res. 2008;14(9):2519–26.CrossRefGoogle Scholar
  6. 6.
    Tsubaki M, Takeda T, Sakamoto K, et al. Bisphosphonates and statins inhibit expression and secretion of MIP-1α via suppression of Ras/MEK/ERK/AML-1A and Ras/PI3K/Akt/AML-1A pathways. Am J Cancer Res. 2014;5(1):168–79.PubMedPubMedCentralGoogle Scholar
  7. 7.
    Tsubaki M, Takeda T, Tomonari Y, et al. The MIP-1α autocrine loop contributes to decreased sensitivity to anticancer drugs. J Cell Physiol. 2018;233(5):4258–71.CrossRefGoogle Scholar
  8. 8.
    Tsubaki M, Mashimo K, Takeda T, et al. Statins inhibited the MIP-1α expression via inhibition of Ras/ERK and Ras/Akt pathways in myeloma cells. Biomed Pharmacother. 2016;78:23–9.CrossRefGoogle Scholar
  9. 9.
    Tsubaki M, Komai M, Itoh T, et al. Inhibition of the tumour necrosis factor-alpha autocrine loop enhances the sensitivity of multiple myeloma cells to anticancer drugs. Eur J Cancer. 2013;49(17):3708–17.CrossRefGoogle Scholar
  10. 10.
    Tsubaki M, Kato C, Nishinobo M, et al. Nitrogen-containing bisphosphonate, YM529/ONO-5920, inhibits macrophage inflammatory protein 1 alpha expression and secretion in mouse myeloma cells. Cancer Sci. 2008;99(1):152–8.PubMedGoogle Scholar
  11. 11.
    Tsubaki M, Kato C, Manno M, et al. Macrophage inflammatory protein-1alpha (MIP-1alpha) enhances a receptor activator of nuclear factor kappaB ligand (RANKL) expression in mouse bone marrow stromal cells and osteoblasts through MAPK and PI3 K/Akt pathways. Mol Cell Biochem. 2007;304(1–2):53–60.CrossRefGoogle Scholar
  12. 12.
    Abdi J, Chen G, Chang H. Drug resistance in multiple myeloma: latest findings and new concepts on molecular mechanisms. Oncotarget. 2013;4(12):2186–207.CrossRefGoogle Scholar
  13. 13.
    Tsubaki M, Takeda T, Ogawa N, et al. Overexpression of survivin via activation of ERK1/2, Akt, and NF-κB plays a central role in vincristine resistance in multiple myeloma cells. Leuk Res. 2015;39(4):445–52.CrossRefGoogle Scholar
  14. 14.
    Tsubaki M, Komai M, Itoh T, et al. By inhibiting Src, verapamil and dasatinib overcome multidrug resistance via increased expression of Bim and decreased expressions of MDR1 and survivin in human multidrug-resistant myeloma cells. Leuk Res. 2014;38(1):121–30.CrossRefGoogle Scholar
  15. 15.
    Furukawa Y, Kikuchi J. Epigenetic mechanisms of cell adhesion-mediated drug resistance in multiple myeloma. Int J Hematol. 2016;104(3):281–92.CrossRefGoogle Scholar
  16. 16.
    Sung B, Cho SG, Liu M, et al. Butein, a tetrahydroxychalcone, suppresses cancer-induced osteoclastogenesis through inhibition of receptor activator of nuclear factor-kappaB ligand signaling. Int J Cancer. 2011;129(9):2062–72.CrossRefGoogle Scholar
  17. 17.
    Sampaio MS, Vettore AL, Yamamoto M, et al. Expression of eight genes of nuclear factor-kappa B pathway in multiple myeloma using bone marrow aspirates obtained at diagnosis. Histol Histopathol. 2009;24(8):991–7.PubMedGoogle Scholar
  18. 18.
    Nishida S, Tsubaki M, Hoshino M, et al. Nitrogen-containing bisphosphonate, YM529/ONO-5920 (a novel minodronic acid), inhibits RANKL expression in a cultured bone marrow stromal cell line ST2. Biochem Biophys Res Commun. 2005;328(1):91–7.CrossRefGoogle Scholar
  19. 19.
    Tsubaki M, Kato C, Isono A, et al. Macrophage inflammatory protein-1α induces osteoclast formation by activation of the MEK/ERK/c-Fos pathway and inhibition of the p38MAPK/IRF-3/IFN-β pathway. J Cell Biochem. 2010;111(6):1661–72.CrossRefGoogle Scholar
  20. 20.
    Tsubaki M, Komai M, Itoh T, et al. Nitrogen-containing bisphosphonates inhibit RANKL- and M-CSF-induced osteoclast formation through the inhibition of ERK1/2 and Akt activation. J Biomed Sci. 2014;21:10.CrossRefGoogle Scholar
  21. 21.
    Tsubaki M, Satou T, Itoh T, et al. Bisphosphonate- and statin-induced enhancement of OPG expression and inhibition of CD9, M-CSF, and RANKL expressions via inhibition of the Ras/MEK/ERK pathway and activation of p38MAPK in mouse bone marrow stromal cell line ST2. Mol Cell Endocrinol. 2012;361(1–2):219–31.CrossRefGoogle Scholar
  22. 22.
    Liu H, Tamashiro S, Baritaki S, et al. TRAF6 activation in multiple myeloma: a potential therapeutic target. Clin Lymphoma Myeloma Leuk. 2012;12(3):155–63.CrossRefGoogle Scholar
  23. 23.
    Tsubaki M, Takeda T, Yoshizumi M, et al. RANK-RANKL interactions are involved in cell adhesion-mediated drug resistance in multiple myeloma cell lines. Tumour Biol. 2016;37(7):9099–110.CrossRefGoogle Scholar
  24. 24.
    Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003;423(6937):337–42.CrossRefGoogle Scholar
  25. 25.
    Frame MC. Src in cancer: deregulation and consequences for cell behaviour. Biochim Biophys Acta. 2002;1602(2):114–30.PubMedGoogle Scholar
  26. 26.
    Lin L, Yan F, Zhao D, et al. Reelin promotes the adhesion and drug resistance of multiple myeloma cells via integrin β1 signaling and STAT3. Oncotarget. 2016;7(9):9844–58.PubMedPubMedCentralGoogle Scholar
  27. 27.
    Zheng Y, Yang J, Qian J, et al. PSGL-1/selectin and ICAM-1/CD18 interactions are involved in macrophage-induced drug resistance in myeloma. Leukemia. 2013;27(3):702–10.CrossRefGoogle Scholar
  28. 28.
    Kanda R, Kawahara A, Watari K, et al. Erlotinib resistance in lung cancer cells mediated by integrin β1/Src/Akt-driven bypass signaling. Cancer Res. 2013;73(20):6243–53.CrossRefGoogle Scholar
  29. 29.
    Wu ZH, Lin C, Liu MM, et al. Src inhibition can synergize with gemcitabine and reverse resistance in triple negative Breast cancer cells via the AKT/c-Jun pathway. PLoS ONE. 2016;11(12):e0169230.CrossRefGoogle Scholar
  30. 30.
    Nam HJ, Im SA, Oh DY, et al. Antitumor activity of saracatinib (AZD0530), a c-Src/Abl kinase inhibitor, alone or in combination with chemotherapeutic agents in gastric cancer. Mol Cancer Ther. 2013;12(1):16–26.CrossRefGoogle Scholar
  31. 31.
    Ferreira PA, Ruela-de-Sousa RR, Queiroz KC, et al. Knocking down low molecular weight protein tyrosine phosphatase (LMW-PTP) reverts chemoresistance through inactivation of Src and Bcr-Abl proteins. PLoS ONE. 2012;7(9):e44312.CrossRefGoogle Scholar
  32. 32.
    Coluccia AM, Cirulli T, Neri P, et al. Validation of PDGFRbeta and c-Src tyrosine kinases as tumor/vessel targets in patients with multiple myeloma: preclinical efficacy of the novel, orally available inhibitor dasatinib. Blood. 2008;112(4):1346–56.CrossRefGoogle Scholar
  33. 33.
    de Queiroz Crusoe E, Maiso P, Fernandez-Lazaro D, et al. Transcriptomic rationale for the synergy observed with dasatinib + bortezomib + dexamethasone in multiple myeloma. Ann Hematol. 2012;91(2):257–69.CrossRefGoogle Scholar
  34. 34.
    Ishikawa H, Tsuyama N, Abroun S, et al. Requirements of src family kinase activity associated with CD45 for myeloma cell proliferation by interleukin-6. Blood. 2002;99(6):2172–8.CrossRefGoogle Scholar
  35. 35.
    Ishikawa H, Tsuyama N, Abroun S, et al. Interleukin-6, CD45 and the src-kinases in myeloma cell proliferation. Leuk Lymphoma. 2003;44(9):1477–81.CrossRefGoogle Scholar
  36. 36.
    Wildes TM, Procknow E, Gao F, et al. Dasatinib in relapsed or plateau-phase multiple myeloma. Leuk Lymphoma. 2009;50(1):137–40.CrossRefGoogle Scholar
  37. 37.
    Futosi K, Németh T, Pick R, et al. Dasatinib inhibits proinflammatory functions of mature human neutrophils. Blood. 2012;119(21):4981–91.CrossRefGoogle Scholar
  38. 38.
    Aplenc R, Blaney SM, Strauss LC, et al. Pediatric phase I trial and pharmacokinetic study of dasatinib: a report from the children’s oncology group phase I consortium. J Clin Oncol. 2011;29(7):839–44.CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  1. 1.Division of Pharmacotherapy, Faculty of PharmacyKindai UniversityHigashi-OsakaJapan
  2. 2.Department of PharmacyJapanese Red Cross Society Wakayama Medical CenterWakayamaJapan
  3. 3.Department of Surgery, Faculty of MedicineKindai UniversityOsakasayamaJapan
  4. 4.Department of Pathology, Faculty of MedicineKindai UniversityOsakasayamaJapan

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