Abstract
Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) such as gefitinib and osimertinib have primarily been used as first-line treatments for patients with EGFR-activating mutations in non-small cell lung cancer (NSCLC). Novel biomarkers are required to distinguish patients with lung cancer who are resistant to EGFR-TKIs. The aim of the study is to investigate the expression and functional role of YES1, one of the Src-family kinases, in EGFR-TKI-resistant NSCLC. YES1 expression was elevated in gefitinib-resistant HCC827 (HCC827/GR) cells, harboring EGFR mutations. Moreover, HCC827/GR cells exhibited increased reactive oxygen species (ROS) levels compared to those of the parent cells, resulting in the phosphorylation/activation of YES1 due to oxidation of the cysteine residue. HCC827/GR cells showed elevated expression levels of YES1-associated protein 1 (YAP1), NF-E2-related factor 2 (Nrf2), cancer stemness-related markers, and antioxidant proteins compared to those of the parent cells. Knockdown of YES1 in HCC827/GR cells suppressed YAP1 phosphorylation, leading to the inhibition of Bcl-2, Bcl-xL, and Cyclin D1 expression. Silencing YES1 markedly attenuated the proliferation, migration, and tumorigenicity of HCC827/GR cells. Dasatinib inhibited the proliferation of HCC827/GR cells by targeting YES1-mediated signaling pathways. Furthermore, the combination of gefitinib and dasatinib demonstrated a synergistic effect in suppressing the proliferation of HCC827/GR cells. Notably, YES1- and Nrf2-regulated genes showed a positive regulatory relationship in patients with lung cancer and in TKI-resistant NSCLC cell lines. Taken together, these findings suggest that modulation of YES1 expression and activity may be an attractive therapeutic strategy for the treatment of drug-resistant NSCLC.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
All data supporting the findings of this study are available within the paper and its Supplementary Information. Raw data are available from the corresponding author upon reasonable request.
References
Biscardi JS, Maa MC, Tice DA et al (1999) c-Src-mediated phosphorylation of the epidermal growth factor receptor on Tyr845 and Tyr1101 is associated with modulation of receptor function. J Biol Chem 274(12):8335–8343. https://doi.org/10.1074/jbc.274.12.8335
Cheong HT, Xu F, Choy CT et al (2018) Upregulation of Bcl2 in NSCLC with acquired resistance to EGFR-TKI. Oncol Lett 15(1):901–907. https://doi.org/10.3892/ol.2017.7377
Ciamporcero E, Daga M, Pizzimenti S et al (2018) Crosstalk between Nrf2 and YAP contributes to maintaining the antioxidant potential and chemoresistance in bladder cancer. Free Radic Biol Med 115:447–457. https://doi.org/10.1016/j.freeradbiomed.2017.12.005
Ehmer U, Sage J (2016) Control of proliferation and cancer growth by the Hippo signaling pathway. Mol Cancer Res 14(2):127–140. https://doi.org/10.1158/1541-7786.MCR-15-0305
Fan PD, Narzisi G, Jayaprakash AD et al (2018) YES1 amplification is a mechanism of acquired resistance to EGFR inhibitors identified by transposon mutagenesis and clinical genomics. Proc Natl Acad Sci U S A 115(26):E6030–E6038. https://doi.org/10.1073/pnas.1717782115
Formisano L, Nappi L, Rosa R et al (2014) Epidermal growth factor-receptor activation modulates Src-dependent resistance to lapatinib in breast cancer models. Breast Cancer Res 16(3):R45. https://doi.org/10.1186/bcr3650
Garmendia I, Pajares MJ, Hermida-Prado F et al (2019) YES1 drives lung cancer growth and progression and predicts sensitivity to dasatinib. Am J Respir Crit Care Med 200(7):888–899. https://doi.org/10.1164/rccm.201807-1292OC
Giannoni E, Buricchi F, Raugei G et al (2005) Intracellular reactive oxygen species activate Src tyrosine kinase during cell adhesion and anchorage-dependent cell growth. Mol Cell Biol 25(15):6391–6403. https://doi.org/10.1128/MCB.25.15.6391-6403.2005
Griffin R, Ramirez RA (2017) Molecular targets in non-small cell lung cancer. Ochsner J 17(4):388–392
Hamanaka N, Nakanishi Y, Mizuno T et al (2019) YES1 is a targetable oncogene in cancers harboring YES1 gene amplification. Cancer Res 79(22):5734–5745. https://doi.org/10.1158/0008-5472.CAN-18-3376
Han S, Meng Y, Tong Q et al (2014) The ErbB2-targeting antibody trastuzumab and the small-molecule SRC inhibitor saracatinib synergistically inhibit ErbB2-overexpressing gastric cancer. Mabs 6(2):403–408. https://doi.org/10.4161/mabs.27443
Heppner DE (2021) Structural insights into redox-active cysteine residues of the Src family kinases. Redox Biol 41:101934. https://doi.org/10.1016/j.redox.2021.101934
Hong AW, Meng Z, Yuan HX et al (2017) Osmotic stress-induced phosphorylation by NLK at Ser128 activates YAP. EMBO Rep 18(1):72–86. https://doi.org/10.15252/embr.201642681
Hsu SC, Miller SA, Wang Y et al (2009) Nuclear EGFR is required for cisplatin resistance and DNA repair. Am J Transl Res 1(3):249–258
Hsu WH, Yang JC, Mok TS et al (2018) Overview of current systemic management of EGFR-mutant NSCLC. Ann Oncol 29(suppl_1):i3–i9. https://doi.org/10.1093/annonc/mdx702
Huang WC, Chen YJ, Li LY et al (2011) Nuclear translocation of epidermal growth factor receptor by Akt-dependent phosphorylation enhances breast cancer-resistant protein expression in gefitinib-resistant cells. J Biol Chem 286(23):20558–20568. https://doi.org/10.1074/jbc.M111.240796
Huang JC, Yue ZP, Yu HF et al (2022) TAZ ameliorates the microglia-mediated inflammatory response via the Nrf2-ROS-NF-κB pathway. Mol Ther Nucleic Acids 28:435–449. https://doi.org/10.1016/j.omtn.2022.03.025
Ichihara E, Westover D, Meador CB et al (2017) SFK/FAK signaling attenuates osimertinib efficacy in both drug-sensitive and drug-resistant models of EGFR-mutant lung cancer. Cancer Res 77(11):2990–3000. https://doi.org/10.1158/0008-5472.CAN-16-2300
Iida M, Brand TM, Campbell DA et al (2013) Yes and Lyn play a role in nuclear translocation of the epidermal growth factor receptor. Oncogene 32(6):759–767. https://doi.org/10.1038/onc.2012.90
Jin H, Jiang AY, Wang H et al (2017) Dihydroartemisinin and gefitinib synergistically inhibit NSCLC cell growth and promote apoptosis via the Akt/mTOR/STAT3 pathway. Mol Med Rep 16(3):3475–3481. https://doi.org/10.3892/mmr.2017.6989
Kahroba H, Shirmohamadi M, Hejazi MS et al (2019) The role of Nrf2 signaling in cancer stem cells: from stemness and self-renewal to tumorigenesis and chemoresistance. Life Sci 239:116986. https://doi.org/10.1016/j.lfs.2019.116986
Kemble DJ, Sun G (2009) Direct and specific inactivation of protein tyrosine kinases in the Src and FGFR families by reversible cysteine oxidation. Proc Natl Acad Sci U S A 106(13):5070–5075. https://doi.org/10.1073/pnas.0806117106
Lamar JM, Stern P, Liu H et al (2012) The Hippo pathway target, YAP, promotes metastasis through its TEAD-interaction domain. Proc Natl Acad Sci U S A 109(37):E2441–E2450. https://doi.org/10.1073/pnas.1212021109
Levy D, Adamovich Y, Reuven N et al (2008) Yap1 phosphorylation by c-Abl is a critical step in selective activation of proapoptotic genes in response to DNA damage. Mol Cell 29(3):350–361. https://doi.org/10.1016/j.molcel.2007.12.022
Li C, Iida M, Dunn EF et al (2009) Nuclear EGFR contributes to acquired resistance to cetuximab. Oncogene 28(43):3801–3813. https://doi.org/10.1038/onc.2009.234
Lin JJ, Shaw AT (2016) Resisting resistance: Targeted therapies in lung cancer. Trends Cancer 2(7):350–364. https://doi.org/10.1016/j.trecan.2016.05.010
Lin YT, Chen JS, Liao WY et al (2019) Clinical outcomes and secondary epidermal growth factor receptor (EGFR) T790M mutation among first-line gefitinib, erlotinib and afatinib-treated non-small cell lung cancer patients with activating EGFR mutations. Int J Cancer 144(11):2887–2896. https://doi.org/10.1002/ijc.32025
Liu Z, Gao W (2019) Overcoming acquired resistance of gefitinib in lung cancer cells without T790M by AZD9291 or Twist1 knockdown in vitro and in vivo. Arch Toxicol 93(6):1555–1571. https://doi.org/10.1007/s00204-019-02453-2
Lo Sardo F, Strano S, Blandino G (2018) YAP and TAZ in lung cancer: oncogenic role and clinical targeting. Cancers (basel) 10(5):137. https://doi.org/10.3390/cancers10050137
Ma CS, Lv QM, Zhang KR et al (2021) NRF2-GPX4/SOD2 axis imparts resistance to EGFR-tyrosine kinase inhibitors in non-small-cell lung cancer cells. Acta Pharmacol Sin 42(4):613–623. https://doi.org/10.1038/s41401-020-0443-1
Maa MC, Leu TH, McCarley DJ et al (1995) Potentiation of epidermal growth factor receptor-mediated oncogenesis by c-Src: implications for the etiology of multiple human cancers. Proc Natl Acad Sci U S A 92(15):6981–6985. https://doi.org/10.1073/pnas.92.15.6981
Mayor S (2017) Osimertinib effective in EGFR T790M-positive lung cancer. Lancet Oncol 18(1):e9. https://doi.org/10.1016/S1470-2045(16)30654-4
Minari R, Valentini S, Madeddu D et al (2022) YES1 and MYC amplifications as synergistic resistance mechanisms to different generation ALK tyrosine kinase inhibitors in advanced NSCLC: brief report of clinical and preclinical proofs. JTO Clin Res Rep 3(2):100278. https://doi.org/10.1016/j.jtocrr.2022.100278
Morin MJ (2000) From oncogene to drug: development of small molecule tyrosine kinase inhibitors as anti-tumor and anti-angiogenic agents. Oncogene 19(56):6574–6583. https://doi.org/10.1038/sj.onc.1204102
Ortiz MA, Mikhailova T, Li X et al (2021) Src family kinases, adaptor proteins and the actin cytoskeleton in epithelial-to-mesenchymal transition. Cell Commun Signal 19(1):67. https://doi.org/10.1186/s12964-021-00750-x
Peng W, Yao C, Pan Q et al (2023) Novel considerations on EGFR-based therapy as a contributor to cancer cell death in NSCLC. Front Oncol 13:1120278. https://doi.org/10.3389/fonc.2023.1120278
Rao S, Gurbani D, Du G et al (2019) Leveraging compound promiscuity to identify targetable cysteines within the kinome. Cell Chem Biol 26(6):818–829. https://doi.org/10.1016/j.chembiol.2019.02.021
Rho JK, Choi YJ, Kim SY et al (2014) MET and AXL inhibitor NPS-1034 exerts efficacy against lung cancer cells resistant to EGFR kinase inhibitors because of MET or AXL activation. Cancer Res 74(1):253–262. https://doi.org/10.1158/0008-5472.CAN-13-1103
Rosenbluh J, Nijhawan D, Cox AG et al (2012) beta-Catenin-driven cancers require a YAP1 transcriptional complex for survival and tumorigenesis. Cell 151(7):1457–1473. https://doi.org/10.1016/j.cell.2012.11.026
Roskoski R Jr (2015) Src protein-tyrosine kinase structure, mechanism, and small molecule inhibitors. Pharmacol Res 94:9–25. https://doi.org/10.1016/j.phrs.2015.01.003
Rotow J, Bivona TG (2017) Understanding and targeting resistance mechanisms in NSCLC. Nat Rev Cancer 17(11):637–658. https://doi.org/10.1038/nrc.2017.84
Rudin CM, Brambilla E, Faivre-Finn C et al (2021) Small-cell lung cancer. Nat Rev Dis Primers 7(1):3. https://doi.org/10.1038/s41572-020-00235-0
Sanchez-Ortega M, Carrera AC, Garrido A (2021) Role of NRF2 in lung cancer. Cells 10(8):1879. https://doi.org/10.3390/cells10081879
Sato H, Kubota D, Qiao H et al (2022) SRC family kinase inhibition targets YES1 and YAP1 as primary drivers of lung cancer and as mediators of acquired resistance to ALK and epidermal growth factor receptor inhibitors. JCO Precis Oncol 6:e2200088. https://doi.org/10.1200/PO.22.00088
Shah R, Lester JF (2020) Tyrosine kinase inhibitors for the treatment of EGFR mutation-positive non-small-cell lung cancer: a clash of the generations. Clin Lung Cancer 21(3):e216–e228. https://doi.org/10.1016/j.cllc.2019.12.003
Siegel RL, Miller KD, Wagle NS et al (2023) (2023) Cancer statistics. CA Cancer J Clin 73(1):17–48. https://doi.org/10.3322/caac.21763
Song J, Zhu J, Zhao Q et al (2015) Gefitinib causes growth arrest and inhibition of metastasis in human chondrosarcoma cells. J BUON 20(3):894–901
Takeda T, Yamamoto H, Kanzaki H et al (2017) Yes1 signaling mediates the resistance to Trastuzumab/Lap atinib in breast cancer. PLoS ONE 12(2):e0171356. https://doi.org/10.1371/journal.pone.0171356
Tamm C, Bower N, Anneren C (2011) Regulation of mouse embryonic stem cell self-renewal by a Yes-YAP-TEAD2 signaling pathway downstream of LIF. J Cell Sci 124(Pt 7):1136–1144. https://doi.org/10.1242/jcs.075796
Tian X, Gu T, Lee MH et al (1877) (2022) Challenge and countermeasures for EGFR targeted therapy in non-small cell lung cancer. Biochim Biophys Acta Rev Cancer 1877:188645. https://doi.org/10.1016/j.bbcan.2021.188645
Tice DA, Biscardi JS, Nickles AL et al (1999) Mechanism of biological synergy between cellular Src and epidermal growth factor receptor. Proc Natl Acad Sci U S A 96(4):1415–1420. https://doi.org/10.1073/pnas.96.4.1415
Wheeler DL, Iida M, Kruser TJ et al (2009) Epidermal growth factor receptor cooperates with Src family kinases in acquired resistance to cetuximab. Cancer Biol Ther 8(8):696–703. https://doi.org/10.4161/cbt.8.8.7903
Xu W, Harrison SC, Eck MJ (1997) Three-dimensional structure of the tyrosine kinase c-Src. Nature 385(6617):595–602. https://doi.org/10.1038/385595a0
Zaidi SK, Sullivan AJ, Medina R et al (2004) Tyrosine phosphorylation controls Runx2-mediated subnuclear targeting of YAP to repress transcription. EMBO J 23(4):790–799. https://doi.org/10.1038/sj.emboj.7600073
Zanconato F, Cordenonsi M, Piccolo S (2016) YAP/TAZ at the roots of cancer. Cancer Cell 29(6):783–803. https://doi.org/10.1016/j.ccell.2016.05.005
Zhang J, Kalyankrishna S, Wislez M et al (2007) SRC-family kinases are activated in non-small cell lung cancer and promote the survival of epidermal growth factor receptor-dependent cell lines. Am J Pathol 170(1):366–376. https://doi.org/10.2353/ajpath.2007.060706
Zhao B, Wei X, Li W et al (2007) Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control. Genes Dev 21(21):2747–2761. https://doi.org/10.1101/gad.1602907
Acknowledgements
This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2020R1A2C1103139 to D.-H.K. and NRF-2021R1C1C1012378 to J.L.). We would like to sincerely thank Prof. Jin Kyung Rho for providing us access to the gefitinib-resistant HCC827 cells.
Author information
Authors and Affiliations
Contributions
DHK conceived and designed the experiments. EJK performed all experiments. EJK, JL, and DHK collected and analyzed the data. EJK, JL, and DHK performed statistical analyses, interpreted the experimental results, and prepared the figures. All the authors have read and approved the final version of the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Kook, E., Lee, J. & Kim, DH. YES1 as a potential target to overcome drug resistance in EGFR-deregulated non-small cell lung cancer. Arch Toxicol 98, 1437–1455 (2024). https://doi.org/10.1007/s00204-024-03693-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-024-03693-7