Vorinostat enhances gefitinib‑induced cell death through reactive oxygen species‑dependent cleavage of HSP90 and its clients in non‑small cell lung cancer with the EGFR mutation

Park,Sang  Eun; Kim,Dong  Eun; Kim,Mi   Joung; Lee,Jee   Suk; Rho,Jin   Kyung; Jeong,Seong‑Yun; Choi,Eun   Kyung; Kim,Choung‑Soo; Hwang,Jung   Jin

doi:10.3892/or.2018.6814

Vorinostat enhances gefitinib‑induced cell death through reactive oxygen species‑dependent cleavage of HSP90 and its clients in non‑small cell lung cancer with the EGFR mutation

Authors:
- Sang Eun Park
- Dong Eun Kim
- Mi Joung Kim
- Jee Suk Lee
- Jin Kyung Rho
- Seong‑Yun Jeong
- Eun Kyung Choi
- Choung‑Soo Kim
- Jung Jin Hwang
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Affiliations: Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea, Department of Radiation Oncology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea
Published online on: October 22, 2018 https://doi.org/10.3892/or.2018.6814
Pages: 525-533

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Abstract

Although different mechanisms of acquired resistance to epidermal growth factor receptor (EGFR)‑tyrosine kinase inhibitors (TKIs) have been reported in non‑small cell lung cancers (NSCLCs), the optimal treatment for patients with acquired resistance has not been clearly defined. The purpose of this study was to investigate the antitumor effects of gefitinib in combination with vorinostat, a potent histone deacetylase inhibitor (HDACI), and their associated molecular mechanisms in relation to activating apoptosis in NSCLC. The treatment using a combination of vorinostat and gefitinib was more potent in promoting cell death by activating apoptosis than gefitinib alone in parental PC9 cells that harbor an EGFR‑activating mutation (EGFR exon 19 deletion) and gefitinib‑resistant PC9 cells (PC9GR) with an EGFR T790M mutation. This combination induced heat shock protein 90 (HSP90) cleavage and reduced the level of HSP90 client proteins, including EGFR, MET and AKT, in PC9 and PC9GR cells. The addition of 4‑(2‑aminoethyl) benzenesulfonyl fluoride hydrochloride, a scavenger of reactive oxygen species (ROS), inhibited the degradation of HSP90 client proteins and HSP90 cleavage that was induced by co‑treatment as well as the cleavage of caspase‑3, caspase‑8, and caspase‑9 and cell death. We also observed that cleavage of HSP90 and its clients were blocked when caspases were inhibited. These results revealed that co‑treatment with vorinostat and gefitinib induced ROS‑dependent caspase activation, leading to the downregulation of HSP90 clients through HSP90 cleavage. Collectively, our findings provide a new basis for strategies that combine vorinostat with an EGFR‑TKI to reverse EGFR‑TKI resistance in NSCLC.

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1	Beere HM: ‘The stress of dying’: The role of heat shock proteins in the regulation of apoptosis. J Cell Sci. 117:2641–2651. 2004. View Article : Google Scholar : PubMed/NCBI
2	Edwards A, Li J, Atadja P, Bhalla K and Haura EB: Effect of the histone deacetylase inhibitor LBH589 against epidermal growth factor receptor-dependent human lung cancer cells. Mol Cancer Ther. 6:2515–2524. 2007. View Article : Google Scholar : PubMed/NCBI
3	Socinski MA, Goldman J, El-Hariry I, Koczywas M, Vukovic V, Horn L, Paschold E, Salgia R, West H, Sequist LV, et al: A multicenter phase II study of ganetespib monotherapy in patients with genotypically defined advanced non-small cell lung cancer. Clin Cancer Res. 19:3068–3077. 2013. View Article : Google Scholar : PubMed/NCBI
4	Jhaveri K, Taldone T, Modi S and Chiosis G: Advances in the clinical development of heat shock protein 90 (Hsp90) inhibitors in cancers. Biochim Biophys Acta. 1823:742–755. 2012. View Article : Google Scholar : PubMed/NCBI
5	Witta SE, Gemmill RM, Hirsch FR, Coldren CD, Hedman K, Ravdel L, Helfrich B, Dziadziuszko R, Chan DC, Sugita M, et al: Restoring E-cadherin expression increases sensitivity to epidermal growth factor receptor inhibitors in lung cancer cell lines. Cancer Res. 66:944–950. 2006. View Article : Google Scholar : PubMed/NCBI
6	Neal JW and Sequist LV: Complex role of histone deacetylase inhibitors in the treatment of non-small-cell lung cancer. J Clin Oncol. 30:2280–2282. 2012. View Article : Google Scholar : PubMed/NCBI
7	Hoang T, Campbell TC, Zhang C, Kim K, Kolesar JM, Oettel KR, Blank JH, Robinson EG, Ahuja HG, Kirschling RJ, et al: Vorinostat and bortezomib as third-line therapy in patients with advanced non-small cell lung cancer: A wisconsin oncology network phase II study. Invest New Drugs. 32:195–199. 2014. View Article : Google Scholar : PubMed/NCBI
8	Rho JK, Choi YJ, Lee JK, Ryoo BY, Na II, Yang SH, Lee SS, Kim CH, Yoo YD and Lee JC: The role of MET activation in determining the sensitivity to epidermal growth factor receptor tyrosine kinase inhibitors. Mol Cancer Res. 7:1736–1743. 2009. View Article : Google Scholar : PubMed/NCBI
9	Rho JK, Lee IY, Choi YJ, Choi CM, Hur JY, Koh JS, Lee J, Suh BC, Song HJ, Salgaonkar P, et al: Superior efficacy and selectivity of novel small-molecule kinase inhibitors of T790M-mutant EGFR in preclinical models of lung cancer. Cancer Res. 77:1200–1211. 2017. View Article : Google Scholar : PubMed/NCBI
10	Gazdar AF: Epidermal growth factor receptor inhibition in lung cancer: The evolving role of individualized therapy. Cancer Metastasis Rev. 29:37–48. 2010. View Article : Google Scholar : PubMed/NCBI
11	Sadiq AA and Salgia R: MET as a possible target for non-small-cell lung cancer. J Clin Oncol. 31:1089–1096. 2013. View Article : Google Scholar : PubMed/NCBI
12	Mahalingam D, Swords R, Carew JS, Nawrocki ST, Bhalla K and Giles FJ: Targeting HSP90 for cancer therapy. Br J Cancer. 100:1523–1529. 2009. View Article : Google Scholar : PubMed/NCBI
13	Beck R, Verrax J, Gonze T, Zappone M, Pedrosa RC, Taper H, Feron O and Calderon PB: Hsp90 cleavage by an oxidative stress leads to its client proteins degradation and cancer cell death. Biochem Pharmacol. 77:375–383. 2009. View Article : Google Scholar : PubMed/NCBI
14	Pantano C, Shrivastava P, McElhinney B and Janssen-Heininger Y: Hydrogen peroxide signaling through tumor necrosis factor receptor 1 leads to selective activation of c-Jun N-terminal kinase. J Biol Chem. 278:44091–44096. 2003. View Article : Google Scholar : PubMed/NCBI
15	Chen H, Xia Y, Fang D, Hawke D and Lu Z: Caspase-10-mediated heat shock protein 90 beta cleavage promotes UVB irradiation-induced cell apoptosis. Mol Cell Biol. 29:3657–3664. 2009. View Article : Google Scholar : PubMed/NCBI
16	Jackson SE and Chester JD: Personalised cancer medicine. Int J Cancer. 137:262–266. 2015. View Article : Google Scholar : PubMed/NCBI
17	Chong CR and Jänne PA: The quest to overcome resistance to EGFR-targeted therapies in cancer. Nat Med. 19:1389–1400. 2013. View Article : Google Scholar : PubMed/NCBI
18	Wheeler DL, Dunn EF and Harari PM: Understanding resistance to EGFR inhibitors-impact on future treatment strategies. Nat Rev Clin Oncol. 7:493–507. 2010. View Article : Google Scholar : PubMed/NCBI
19	Bose P, Dai Y and Grant S: Histone deacetylase inhibitor (HDACI) mechanisms of action: Emerging insights. Pharmacol Ther. 143:323–336. 2014. View Article : Google Scholar : PubMed/NCBI
20	Minucci S and Pelicci PG: Histone deacetylase inhibitors and the promise of epigenetic (and more) treatments for cancer. Nat Rev Cancer. 6:38–51. 2006. View Article : Google Scholar : PubMed/NCBI
21	Nakagawa T, Takeuchi S, Yamada T, Ebi H, Sano T, Nanjo S, Ishikawa D, Sato M, Hasegawa Y, Sekido Y and Yano S: EGFR-TKI resistance due to BIM polymorphism can be circumvented in combination with HDAC inhibition. Cancer Res. 73:2428–2434. 2013. View Article : Google Scholar : PubMed/NCBI
22	Busser B, Sancey L, Josserand V, Niang C, Khochbin S, Favrot MC, Coll JL and Hurbin A: Amphiregulin promotes resistance to gefitinib in nonsmall cell lung cancer cells by regulating Ku70 acetylation. Mol Ther. 18:536–543. 2010. View Article : Google Scholar : PubMed/NCBI
23	Jeannot V, Busser B, Vanwonterghem L, Michallet S, Ferroudj S, Cokol M, Coll JL, Ozturk M and Hurbin A: Synergistic activity of vorinostat combined with gefitinib but not with sorafenib in mutant KRAS human non-small cell lung cancers and hepatocarcinoma. Onco Targets Ther. 9:6843–6855. 2016. View Article : Google Scholar : PubMed/NCBI
24	Leone A, Roca MS, Ciardiello C, Terranova-Barberio M, Vitagliano C, Ciliberto G, Mancini R, Di Gennaro E, Bruzzese F and Budillon A: Vorinostat synergizes with EGFR inhibitors in NSCLC cells by increasing ROS via up-regulation of the major mitochondrial porin VDAC1 and modulation of the c-Myc-NRF2-KEAP1 pathway. Free Radic Biol Med. 89:287–299. 2015. View Article : Google Scholar : PubMed/NCBI
25	Bali P, Pranpat M, Bradner J, Balasis M, Fiskus W, Guo F, Rocha K, Kumaraswamy S, Boyapalle S, Atadja P, et al: Inhibition of histone deacetylase 6 acetylates and disrupts the chaperone function of heat shock protein 90: A novel basis for antileukemia activity of histone deacetylase inhibitors. J Biol Chem. 280:26729–26734. 2005. View Article : Google Scholar : PubMed/NCBI
26	Scroggins BT, Robzyk K, Wang D, Marcu MG, Tsutsumi S, Beebe K, Cotter RJ, Felts S, Toft D, Karnitz L, et al: An acetylation site in the middle domain of Hsp90 regulates chaperone function. Mol Cell. 25:151–159. 2007. View Article : Google Scholar : PubMed/NCBI
27	Nishioka C, Ikezoe T, Yang J, Takeuchi S, Koeffler HP and Yokoyama A: M S-275, a novel histone deacetylase inhibitor with selectivity against HDAC1, induces degradation of FLT3 via inhibition of chaperone function of heat shock protein 90 in AML cells. Leuk Res. 32:1382–1392. 2008. View Article : Google Scholar : PubMed/NCBI
28	Ruefli AA, Ausserlechner MJ, Bernhard D, Sutton VR, Tainton KM, Kofler R, Smyth MJ and Johnstone RW: The histone deacetylase inhibitor and chemotherapeutic agent suberoylanilide hydroxamic acid (SAHA) induces a cell-death pathway characterized by cleavage of Bid and production of reactive oxygen species. Proc Natl Acad Sci USA. 98:10833–10838. 2001. View Article : Google Scholar : PubMed/NCBI
29	Kim DH, Lee J, Kim KN, Kim HJ, Jeung HC, Chung HC and Kwon HJ: Anti-tumor activity of N-hydroxy-7-(2-naphthylthio) heptanomide, a novel histone deacetylase inhibitor. Biochem Biophys Res Commun. 356:233–238. 2007. View Article : Google Scholar : PubMed/NCBI
30	Ng KP, Hillmer AM, Chuah CT, Juan WC, Ko TK, Teo AS, Ariyaratne PN, Takahashi N, Sawada K, Fei Y, et al: A common BIM deletion polymorphism mediates intrinsic resistance and inferior responses to tyrosine kinase inhibitors in cancer. Nat Med. 18:521–528. 2012. View Article : Google Scholar : PubMed/NCBI
31	Han JY, Lee SH, Lee GK, Yun T, Lee YJ, Hwang KH, Kim JY and Kim HT: Phase I/II study of gefitinib (Iressa(^®)) and vorinostat (IVORI) in previously treated patients with advanced non-small cell lung cancer. Cancer Chemother Pharmacol. 75:475–483. 2015. View Article : Google Scholar : PubMed/NCBI