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Y-box binding protein 1 (YB-1) promotes gefitinib resistance in lung adenocarcinoma cells by activating AKT signaling and epithelial–mesenchymal transition through targeting major vault protein (MVP)

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Abstract

Purpose

Gefitinib is a first-line treatment option for epidermal growth factor receptor (EGFR)-mutated lung adenocarcinoma. However, most patients inevitably develop gefitinib resistance. The mechanism underlying this resistance is not fully understood. Y-box binding protein 1 (YB-1) has been reported to play a role in modulating drug sensitivity, but its role in gefitinib resistance is currently unknown. Here, we investigated the role of YB-1 in gefitinib resistance of lung adenocarcinoma.

Methods

We determined the expression of YB-1, epithelial–mesenchymal transition (EMT) and AKT signaling markers, as well as the viability of lung adenocarcinoma cell lines bearing mutant (HCC827, PC-9) or wild-type (H1299) EGFR. We also evaluated PC-9 cell migration and invasion using transwell assays. The clinical importance of YB-1 and major vault protein (MVP) was evaluated using primary lung adenocarcinoma patient samples.

Results

We found that YB-1 was significantly upregulated in gefitinib-resistant lung adenocarcinoma cells compared to gefitinib-sensitive cells. YB-1 augmented gefitinib resistance by activating the AKT pathway and promoting EMT. Decreased migration and invasion was observed upon MVP silencing in YB-1-overexpressing PC-9 cells, as well as restored gefitinib sensitivity. A retrospective analysis of 85 patients with lung adenocarcinoma revealed that YB-1 levels were significantly increased in tyrosine kinase inhibitor (TKI)-resistant patients compared to those in TKI-sensitive patients, indicating that YB-1 may serve as a biomarker to clinically predict acquired gefitinib resistance.

Conclusion

YB-1 activates AKT signaling and promotes EMT at least in part by directly activating MVP. Hence, targeting the YB-1/MVP axis may help to overcome gefitinib resistance in lung adenocarcinoma patients.

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References

  1. R. Siegel, D. Naishadham, A. Jemal, Cancer statistics, 2013. CA Cancer J Clin 63, 11–30 (2013)

    PubMed  Google Scholar 

  2. T.S. Mok, Y.L. Wu, S. Thongprasert, C.H. Yang, D.T. Chu, N. Saijo, P. Sunpaweravong, B. Han, B. Margono, Y. Ichinose, Y. Nishiwaki, Y. Ohe, J.J. Yang, B. Chewaskulyong, H. Jiang, E.L. Duffield, C.L. Watkins, A.A. Armour, M. Fukuoka, Gefitinib or carboplatin-paclitaxel in pulmonary adenocarcinoma. N Engl J Med 361, 947–957 (2009)

    CAS  PubMed  Google Scholar 

  3. G.R. Oxnard, M.E. Arcila, C.S. Sima, G.J. Riely, J. Chmielecki, M.G. Kris, W. Pao, M. Ladanyi, V.A. Miller, Acquired resistance to EGFR tyrosine kinase inhibitors in EGFR-mutant lung cancer: Distinct natural history of patients with tumors harboring the T790M mutation. Clin Cancer Res 17, 1616–1622 (2011)

    CAS  PubMed  Google Scholar 

  4. D. Jackman, W. Pao, G.J. Riely, J.A. Engelman, M.G. Kris, P.A. Janne, T. Lynch, B.E. Johnson, V.A. Miller, Clinical definition of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small-cell lung cancer. J Clin Oncol 28, 357–360 (2010)

    CAS  PubMed  Google Scholar 

  5. G.R. Oxnard, M.E. Arcila, J. Chmielecki, M. Ladanyi, V.A. Miller, W. Pao, New strategies in overcoming acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in lung cancer. Clin Cancer Res 17, 5530–5537 (2011)

    CAS  PubMed  PubMed Central  Google Scholar 

  6. M. Maemondo, A. Inoue, K. Kobayashi, S. Sugawara, S. Oizumi, H. Isobe, A. Gemma, M. Harada, H. Yoshizawa, I. Kinoshita, Y. Fujita, S. Okinaga, H. Hirano, K. Yoshimori, T. Harada, T. Ogura, M. Ando, H. Miyazawa, T. Tanaka, Y. Saijo, K. Hagiwara, S. Morita, T. Nukiwa, Gefitinib or chemotherapy for non-small-cell lung cancer with mutated EGFR. N Engl J Med 362, 2380–2388 (2010)

    CAS  PubMed  Google Scholar 

  7. Y. Shang, X. Cai, D. Fan, Roles of epithelial-mesenchymal transition in cancer drug resistance. Curr Cancer Drug Targets 13, 915–929 (2013)

    CAS  PubMed  Google Scholar 

  8. A. Voulgari, A. Pintzas, Epithelial-mesenchymal transition in cancer metastasis: Mechanisms, markers and strategies to overcome drug resistance in the clinic. Biochim Biophys Acta 1796, 75–90 (2009)

    CAS  PubMed  Google Scholar 

  9. H. Kajiyama, K. Shibata, M. Terauchi, M. Yamashita, K. Ino, A. Nawa, F. Kikkawa, Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells. Int J Oncol 31, 277–283 (2007)

    CAS  PubMed  Google Scholar 

  10. L. Sun, Y. Yao, B. Liu, Z. Lin, L. Lin, M. Yang, W. Zhang, W. Chen, C. Pan, Q. Liu, E. Song, J. Li, MiR-200b and miR-15b regulate chemotherapy-induced epithelial-mesenchymal transition in human tongue cancer cells by targeting BMI1. Oncogene 31, 432–445 (2012)

    CAS  PubMed  Google Scholar 

  11. L. Li, X. Gu, J. Yue, Q. Zhao, D. Lv, H. Chen, L. Xu, Acquisition of EGFR TKI resistance and EMT phenotype is linked with activation of IGF1R/NF-kappaB pathway in EGFR-mutant NSCLC. Oncotarget Acquisition of EGFR TKI resistance and EMT phenotype is linked with activation of IGF1R/NF-kappaB pathway in EGFR-mutant NSCLC. Oncotarget 8, 92240–92253 (2017)

    PubMed  PubMed Central  Google Scholar 

  12. J. Yue, D. Lv, C. Wang, L. Li, Q. Zhao, H. Chen, L. Xu, Epigenetic silencing of miR-483-3p promotes acquired gefitinib resistance and EMT in EGFR-mutant NSCLC by targeting integrin beta3. Oncogene 37, 4300–4312 (2018)

    CAS  PubMed  PubMed Central  Google Scholar 

  13. L.A. Byers, L. Diao, J. Wang, P. Saintigny, L. Girard, M. Peyton, L. Shen, Y. Fan, U. Giri, P.K. Tumula, M.B. Nilsson, J. Gudikote, H. Tran, R.J. Cardnell, D.J. Bearss, S.L. Warner, J.M. Foulks, S.B. Kanner, V. Gandhi, N. Krett, S.T. Rosen, E.S. Kim, R.S. Herbst, G.R. Blumenschein, J.J. Lee, S.M. Lippman, K.K. Ang, G.B. Mills, W.K. Hong, J.N. Weinstein, I.I. Wistuba, K.R. Coombes, J.D. Minna, J.V. Heymach, An epithelial-mesenchymal transition gene signature predicts resistance to EGFR and PI3K inhibitors and identifies Axl as a therapeutic target for overcoming EGFR inhibitor resistance. Clin Cancer Res 19, 279–290 (2013)

  14. H. Uramoto, T. Iwata, T. Onitsuka, H. Shimokawa, T. Hanagiri, T. Oyama, Epithelial-mesenchymal transition in EGFR-TKI acquired resistant lung adenocarcinoma. Anticancer Res 30, 2513–2517 (2010)

    CAS  PubMed  Google Scholar 

  15. Y. Lin, X. Wang, H. Jin, EGFR-TKI resistance in NSCLC patients: Mechanisms and strategies. Am J Cancer Res 4, 411–435 (2014)

    PubMed  PubMed Central  Google Scholar 

  16. S.E. Witta, R.M. Gemmill, F.R. Hirsch, C.D. Coldren, K. Hedman, L. Ravdel, B. Helfrich, R. Dziadziuszko, D.C. Chan, M. Sugita, Z. Chan, A. Baron, W. Franklin, H.A. Drabkin, L. Girard, A.F. Gazdar, J.D. Minna, P.A. Bunn, Restoring E-cadherin expression increases sensitivity to epidermal growth factor receptor inhibitors in lung cancer cell lines. Cancer Res 66, 944–950 (2006)

    CAS  PubMed  Google Scholar 

  17. K. Kohno, H. Izumi, T. Uchiumi, M. Ashizuka, M. Kuwano, The pleiotropic functions of the Y-box-binding protein, YB-1. Bioessays 25, 691–698 (2003)

  18. M. Kuwano, Y. Oda, H. Izumi, S.J. Yang, T. Uchiumi, Y. Iwamoto, M. Toi, T. Fujii, H. Yamana, H. Kinoshita, T. Kamura, M. Tsuneyoshi, K. Yasumoto, K. Kohno, The role of nuclear Y-box binding protein 1 as a global marker in drug resistance. Mol Cancer Ther 3, 1485–1492 (2004)

    CAS  PubMed  Google Scholar 

  19. E. Mylona, S. Melissaris, I. Giannopoulou, I. Theohari, C. Papadimitriou, A. Keramopoulos, L. Nakopoulou, Y-box-binding protein 1 (YB1) in breast carcinomas: Relation to aggressive tumor phenotype and identification of patients at high risk for relapse. Eur J Surg Oncol 40, 289–296 (2014)

    CAS  PubMed  Google Scholar 

  20. T. Guo, S. Zhao, P. Wang, X. Xue, Y. Zhang, M. Yang, N. Li, Z. Li, L. Xu, L. Jiang, L. Zhao, P.C. Ma, R. Rosell, J. Li, C. Gu, YB-1 regulates tumor growth by promoting MACC1/c-met pathway in human lung adenocarcinoma. Oncotarget 8, 48110–48125 (2017)

    PubMed  PubMed Central  Google Scholar 

  21. S. Zhao, Y. Wang, T. Guo, W. Yu, J. Li, Z. Tang, Z. Yu, L. Zhao, Y. Zhang, Z. Wang, P. Wang, Y. Li, F. Li, Z. Sun, Y. Xuan, R. Tang, W.G. Deng, W. Guo, C. Gu, YBX1 regulates tumor growth via CDC25a pathway in human lung adenocarcinoma. Oncotarget 7, 82139–82157 (2016)

    PubMed  PubMed Central  Google Scholar 

  22. M. Kuwano, T. Shibata, K. Watari, M. Ono, Oncogenic Y-box binding protein-1 as an effective therapeutic target in drug-resistant cancer. Cancer Sci 110, 1536–1543 (2019)

    CAS  PubMed  PubMed Central  Google Scholar 

  23. V. Evdokimova, C. Tognon, T. Ng, P. Ruzanov, N. Melnyk, D. Fink, A. Sorokin, L.P. Ovchinnikov, E. Davicioni, T.J. Triche, P.H. Sorensen, Translational activation of snail1 and other developmentally regulated transcription factors by YB-1 promotes an epithelial-mesenchymal transition. Cancer Cell 15, 402–415 (2009)

    CAS  PubMed  Google Scholar 

  24. U. Stein, S. Bergmann, G.L. Scheffer, R.J. Scheper, H.D. Royer, P.M. Schlag, W. Walther, YB-1 facilitates basal and 5-fluorouracil-inducible expression of the human major vault protein (MVP) gene. Oncogene 24, 3606–3618 (2005)

    CAS  PubMed  Google Scholar 

  25. V.A. Kickhoefer, K.S. Rajavel, G.L. Scheffer, W.S. Dalton, R.J. Scheper, L.H. Rome, Vaults are up-regulated in multidrug-resistant cancer cell lines. J Biol Chem 273, 8971–8974 (1998)

    CAS  PubMed  Google Scholar 

  26. K. Park, The role of major vault protein (MVP) in drug resistance. J Control Release 163, 266 (2012)

    CAS  PubMed  Google Scholar 

  27. Y.S. Xiao, D. Zeng, Y.K. Liang, Y. Wu, M.F. Li, Y.Z. Qi, X.L. Wei, W.H. Huang, M. Chen, G.J. Zhang, Major vault protein is a direct target of Notch1 signaling and contributes to chemoresistance in triple-negative breast cancer cells. Cancer Lett 440–441, 156–167 (2019)

  28. X. Miao, Y. Wu, Y. Wang, X. Zhu, H. Yin, Y. He, C. Li, Y. Liu, X. Lu, Y. Chen, R. Shen, X. Xu, S. He, Y-box-binding protein-1 (YB-1) promotes cell proliferation, adhesion and drug resistance in diffuse large B-cell lymphoma. Exp Cell Res 346, 157–166 (2016)

    CAS  PubMed  Google Scholar 

  29. X.H. Kang, Z.Y. Xu, Y.B. Gong, L.F. Wang, Z.Q. Wang, L. Xu, F. Cao, M.J. Liao, Bufalin reverses HGF-induced resistance to EGFR-TKIs in EGFR mutant lung Cancer cells via blockage of met/PI3k/Akt pathway and induction of apoptosis. Evid Based Complement Alternat Med 2013, 243859 (2013)

    PubMed  PubMed Central  Google Scholar 

  30. K. Sakai, A. Horiike, D.L. Irwin, K. Kudo, Y. Fujita, A. Tanimoto, T. Sakatani, R. Saito, K. Kaburaki, N. Yanagitani, F. Ohyanagi, M. Nishio, K. Nishio, Detection of epidermal growth factor receptor T790M mutation in plasma DNA from patients refractory to epidermal growth factor receptor tyrosine kinase inhibitor. Cancer Sci 104, 1198–1204 (2013)

    CAS  PubMed  PubMed Central  Google Scholar 

  31. T. Fujii, G. Yokoyama, H. Takahashi, U. Toh, M. Kage, M. Ono, K. Shirouzu, M. Kuwano, Preclinical and clinical studies of novel breast cancer drugs targeting molecules involved in protein kinase C signaling, the putative metastasis-suppressor gene Cap43 and the Y-box binding protein-1. Curr Med Chem 15, 528–537 (2008)

    CAS  PubMed  Google Scholar 

  32. Y. Oda, Y. Ohishi, Y. Basaki, H. Kobayashi, T. Hirakawa, N. Wake, M. Ono, K. Nishio, M. Kuwano, M. Tsuneyoshi, Prognostic implications of the nuclear localization of Y-box-binding protein-1 and CXCR4 expression in ovarian cancer: Their correlation with activated Akt. LRP/MVP and P-glycoprotein expression Cancer Sci 98, 1020–1026 (2007)

    CAS  PubMed  Google Scholar 

  33. Y. Fujiwara-Okada, Y. Matsumoto, J. Fukushi, N. Setsu, S. Matsuura, S. Kamura, T. Fujiwara, K. Iida, M. Hatano, A. Nabeshima, H. Yamada, M. Ono, Y. Oda, Y. Iwamoto, Y-box binding protein-1 regulates cell proliferation and is associated with clinical outcomes of osteosarcoma. Br J Cancer 108, 836–847 (2013)

    CAS  PubMed  PubMed Central  Google Scholar 

  34. M. Shiota, A. Takeuchi, Y. Song, A. Yokomizo, E. Kashiwagi, T. Uchiumi, K. Kuroiwa, K. Tatsugami, N. Fujimoto, Y. Oda, S. Naito, Y-box binding protein-1 promotes castration-resistant prostate cancer growth via androgen receptor expression. Endocr Relat Cancer 18, 505–517 (2011)

    CAS  PubMed  Google Scholar 

  35. J. Zheng, W. Dong, J. Zhang, G. Li, H. Gong, YB-1, a new biomarker of glioma progression, is associated with the prognosis of glioma patients. Acta Biochim Biophys Sin Shanghai 48, 318–325 (2016)

    CAS  PubMed  PubMed Central  Google Scholar 

  36. A. Maciejczyk, J. Szelachowska, M. Ekiert, R. Matkowski, A. Halon, H. Lage, P. Surowiak, Elevated nuclear YB1 expression is associated with poor survival of patients with early breast cancer. Anticancer Res 32, 3177–3184 (2012)

    PubMed  Google Scholar 

  37. O. Gluz, K. Mengele, M. Schmitt, R. Kates, R. Diallo-Danebrock, F. Neff, H.D. Royer, N. Eckstein, S. Mohrmann, E. Ting, M. Kiechle, C. Poremba, U. Nitz, N. Harbeck, Y-box-binding protein YB-1 identifies high-risk patients with primary breast cancer benefiting from rapidly cycled tandem high-dose adjuvant chemotherapy. J Clin Oncol 27, 6144–6151 (2009)

    CAS  PubMed  Google Scholar 

  38. T. Ohga, K. Koike, M. Ono, Y. Makino, Y. Itagaki, M. Tanimoto, M. Kuwano, K. Kohno, Role of the human Y box-binding protein YB-1 in cellular sensitivity to the DNA-damaging agents cisplatin, mitomycin C, and ultraviolet light. Cancer Res 56, 4224–4228 (1996)

    CAS  PubMed  Google Scholar 

  39. K. Shibahara, T. Uchiumi, T. Fukuda, S. Kura, Y. Tominaga, Y. Maehara, K. Kohno, Y. Nakabeppu, T. Tsuzuki, M. Kuwano, Targeted disruption of one allele of the Y-box binding protein-1 (YB-1) gene in mouse embryonic stem cells and increased sensitivity to cisplatin and mitomycin C. Cancer Sci 95, 348–353 (2004)

    CAS  PubMed  Google Scholar 

  40. A.L. Stratford, G. Habibi, A. Astanehe, H. Jiang, K. Hu, E. Park, A. Shadeo, T.P. Buys, W. Lam, T. Pugh, M. Marra, T.O. Nielsen, U. Klinge, P.R. Mertens, S. Aparicio, S.E. Dunn, Epidermal growth factor receptor (EGFR) is transcriptionally induced by the Y-box binding protein-1 (YB-1) and can be inhibited with Iressa in basal-like breast cancer, providing a potential target for therapy. Breast Cancer Res 9, R61 (2007)

    PubMed  PubMed Central  Google Scholar 

  41. M.A. Izquierdo, G.L. Scheffer, M.J. Flens, G. Giaccone, H.J. Broxterman, C.J. Meijer, P. van der Valk, R.J. Scheper, Broad distribution of the multidrug resistance-related vault lung resistance protein in normal human tissues and tumors. Am J Pathol 148, 877–887 (1996)

    CAS  PubMed  PubMed Central  Google Scholar 

  42. A.C. Siva, S. Raval-Fernandes, A.G. Stephen, M.J. LaFemina, R.J. Scheper, V.A. Kickhoefer, L.H. Rome, Up-regulation of vaults may be necessary but not sufficient for multidrug resistance. Int J Cancer 92, 195–202 (2001)

    CAS  PubMed  Google Scholar 

  43. C. Lange, W. Walther, H. Schwabe, U. Stein, Cloning and initial analysis of the human multidrug resistance-related MVP/LRP gene promoter. Biochem Biophys Res Commun 278, 125–133 (2000)

    CAS  PubMed  Google Scholar 

  44. A. Hyogotani, K. Ito, K. Yoshida, H. Izumi, K. Kohno, J. Amano, Association of nuclear YB-1 localization with lung resistance-related protein and epidermal growth factor receptor expression in lung cancer. Clin Lung Cancer 13, 375–384 (2012)

    CAS  PubMed  Google Scholar 

  45. B. Du, J.S. Shim, Targeting epithelial-Mesenchymal transition (EMT) to overcome drug resistance in cancer. Molecules 21, 965 (2016)

  46. M.E. Goldsmith, M.J. Madden, C.S. Morrow, K.H. Cowan, A Y-box consensus sequence is required for basal expression of the human multidrug resistance (mdr1) gene. J Biol Chem 268, 5856–5860 (1993)

    CAS  PubMed  Google Scholar 

  47. D. Nantajit, D. Lin, J.J. Li, The network of epithelial-mesenchymal transition: Potential new targets for tumor resistance. J Cancer Res Clin Oncol 141, 1697–1713 (2015)

    CAS  PubMed  Google Scholar 

  48. H. Sui, L. Zhu, W. Deng, Q. Li, Epithelial-mesenchymal transition and drug resistance: Role, molecular mechanisms, and therapeutic strategies. Oncol Res Treat 37, 584–589 (2014)

    CAS  PubMed  Google Scholar 

  49. Q.B. She, D. Solit, A. Basso, M.M. Moasser, Resistance to gefitinib in PTEN-null HER-overexpressing tumor cells can be overcome through restoration of PTEN function or pharmacologic modulation of constitutive phosphatidylinositol 3′-kinase/Akt pathway signaling. Clin Cancer Res 9, 4340–4346 (2003)

    CAS  PubMed  Google Scholar 

  50. M. Karimi Roshan, A. Soltani, A. Soleimani, K. Rezaie Kahkhaie, A.R. Afshari, M. Soukhtanloo, Role of AKT and mTOR signaling pathways in the induction of epithelial-mesenchymal transition (EMT) process. Biochimie 165, 229–234 (2019)

    CAS  PubMed  Google Scholar 

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Availability of data and material

The datasets used or analysed during the current study are available from the corresponding author on reasonable request.

Funding

This project was supported by the National Natural Science Foundation of China (Grant No.81502634).

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Author notes

  1. Lei Lou and Juan Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, The Second Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, 050000, People’s Republic of China

    Lei Lou, Juan Wang, Yuehong Li & Xianghong Zhang

  2. Laboratory of Pathology, Hebei Medical University, Shijiazhuang, Hebei Province Shijiazhuang, Hebei, 050017, People’s Republic of China

    Lei Lou, Fengzhu Lv, Lingxiao Xing, Haitao Shen & Xianghong Zhang

  3. Department of Radiotherapy, The Second Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, 050000, People’s Republic of China

    Guohui Wang

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  1. Lei Lou
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Contributions

LL was responsible for the study concept and design. FZ, GW and HS acquired the data for the study. JW and YL were responsible for data analysis and interpretation. LL prepared the manuscript and LX reviewed the manuscript. XZ is corresponding author. All other authors read and approved the final version of the manuscript.

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Correspondence to Xianghong Zhang.

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Lou, L., Wang, J., Lv, F. et al. Y-box binding protein 1 (YB-1) promotes gefitinib resistance in lung adenocarcinoma cells by activating AKT signaling and epithelial–mesenchymal transition through targeting major vault protein (MVP). Cell Oncol. 44, 109–133 (2021). https://doi.org/10.1007/s13402-020-00556-y

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