Interleukin‑6 identified as an important factor in hypoxia‑ and aldehyde dehydrogenase‑based gefitinib adaptive resistance in non‑small cell lung cancer cells

An,Shi‑Min; Lei,Hui‑Min; Ding,Xu‑Ping; Sun,Fan; Zhang,Chun; Tang,Ya‑Bin; Chen,Hong‑Zhuan; Shen,Ying; Zhu,Liang

doi:10.3892/ol.2017.6613

Interleukin‑6 identified as an important factor in hypoxia‑ and aldehyde dehydrogenase‑based gefitinib adaptive resistance in non‑small cell lung cancer cells

Authors:
- Shi‑Min An
- Hui‑Min Lei
- Xu‑Ping Ding
- Fan Sun
- Chun Zhang
- Ya‑Bin Tang
- Hong‑Zhuan Chen
- Ying Shen
- Liang Zhu
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Affiliations: Department of Pharmacology and Chemical Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China
Published online on: July 20, 2017 https://doi.org/10.3892/ol.2017.6613
Pages: 3445-3454
Copyright: © An et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Gefitinib resistance and relapse of the disease were the greatest challenges facing clinical therapy of non‑small‑cell lung cancer (NSCLC). Of note, regarding the hypoxia condition in solid tumor tissues in vivo, roles of hypoxia in gefitinib adaptive resistance and its association with lung cancer stem cells (LCSCs) have not been fully elucidated. In the present study, the role of hypoxia in gefitinib adaptive resistance and its association with aldehyde dehydrogenase (ALDH)‑based LCSC gefitinib resistance were comparatively studied using RNA‑sequencing (RNA‑seq) technology. Co‑treatment of PC9 cells with gefitinib and hypoxia (1% O2) significantly enhanced adaptive resistance compared with gefitinib or hypoxia treatment alone. An ALDEFLUOR assay demonstrated that there was a significant increase of ALDH expression level in hypoxia and gefitinib co‑treated PC9 cells, in addition to a higher ratio of G0/G1 quiescent cell enrichment and acquisition of epithelial‑mesenchymal transition. RNA‑seq analysis revealed that interleukin‑6 (IL‑6) is an important common factor in hypoxia and ALDH‑based gefitinib resistance, supported by inflammation‑associated tumor necrosis factor, nuclear factor‑κB and Janus kinase‑signal transducer and activator of transcription signaling pathway enrichment. Furthermore, exposure of PC9 and HCC827 cells to IL‑6 increased gefitinib adaptive resistance. Consequently, IL‑6 expression level was a poor prognostic marker for patients with NSCLC and adenocarcinoma. Thus, targeting IL‑6 combined with tyrosine kinase inhibitor treatment may be promising in NSCLC clinical therapy in the future.

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1	Herbst RS, Heymach JV and Lippman SM: Lung cancer. N Engl J Med. 359:1367–1380. 2008. View Article : Google Scholar : PubMed/NCBI
2	Jamal-Hanjani M, Wilson GA, McGranahan N, Birkbak NJ, Watkins TBK, Veeriah S, Shafi S, Johnson DH, Mitter R, Rosenthal R, et al: Tracking the evolution of non-small-cell lung cancer. N Engl J Med. 376:2109–2121. 2017. View Article : Google Scholar : PubMed/NCBI
3	Lamouille S, Xu J and Derynck R: Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 15:178–196. 2014. View Article : Google Scholar : PubMed/NCBI
4	Singh A and Settleman J: EMT, cancer stem cells and drug resistance: An emerging axis of evil in the war on cancer. Oncogene. 29:4741–4751. 2010. View Article : Google Scholar : PubMed/NCBI
5	Graves EE, Maity A and Le QT: The tumor microenvironment in non-small-cell lung cancer. Semin Radiat Oncol. 20:156–163. 2010. View Article : Google Scholar : PubMed/NCBI
6	Murakami A, Takahashi F, Nurwidya F, Kobayashi I, Minakata K, Hashimoto M, Nara T, Kato M, Tajima K, Shimada N, et al: Hypoxia increases gefitinib-resistant lung cancer stem cells through the activation of insulin-like growth factor 1 receptor. PLoS One. 9:e864592014. View Article : Google Scholar : PubMed/NCBI
7	Mazumdar J, Dondeti V and Simon MC: Hypoxia-inducible factors in stem cells and cancer. J Cell Mol Med. 13:4319–4328. 2009. View Article : Google Scholar : PubMed/NCBI
8	Iida H, Suzuki M, Goitsuka R and Ueno H: Hypoxia induces CD133 expression in human lung cancer cells by up-regulation of OCT3/4 and SOX2. Int J Oncol. 40:71–79. 2012.PubMed/NCBI
9	Ma I and Allan AL: The role of human aldehyde dehydrogenase in normal and cancer stem cells. Stem Cell Rev. 7:292–306. 2011. View Article : Google Scholar : PubMed/NCBI
10	Győrffy B, Surowiak P, Budczies J and Lánczky A: Online survival analysis software to assess the prognostic value of biomarkers using transcriptomic data in non-small-cell lung cancer. PLoS One. 8:e822412013. View Article : Google Scholar : PubMed/NCBI
11	Curtis SJ, Sinkevicius KW, Li D, Lau AN, Roach RR, Zamponi R, Woolfenden AE, Kirsch DG, Wong KK and Kim CF: Primary tumor genotype is an important determinant in identification of lung cancer propagating cells. Cell Stem Cell. 7:127–133. 2010. View Article : Google Scholar : PubMed/NCBI
12	Freitas DP, Teixeira CA, Santos-Silva F, Vasconcelos MH and Almeida GM: Therapy-induced enrichment of putative lung cancer stem-like cells. Int J Cancer. 134:1270–1278. 2014. View Article : Google Scholar : PubMed/NCBI
13	Mizuno T, Suzuki N, Makino H, Furui T, Morii E, Aoki H, Kunisada T, Yano M, Kuji S, Hirashima Y, et al: Cancer stem-like cells of ovarian clear cell carcinoma are enriched in the ALDH-high population associated with an accelerated scavenging system in reactive oxygen species. Gynecol Oncol. 137:299–305. 2015. View Article : Google Scholar : PubMed/NCBI
14	Brennan SK, Meade B, Wang Q, Merchant AA, Kowalski J and Matsui W: Mantle cell lymphoma activation enhances bortezomib sensitivity. Blood. 116:4185–4191. 2010. View Article : Google Scholar : PubMed/NCBI
15	Terai H, Soejima K, Yasuda H, Nakayama S, Hamamoto J, Arai D, Ishioka K, Ohgino K, Ikemura S, Sato T, et al: Activation of the FGF2-FGFR1 autocrine pathway: A novel mechanism of acquired resistance to gefitinib in NSCLC. Mol Cancer Res. 11:759–767. 2013. View Article : Google Scholar : PubMed/NCBI
16	Ware KE, Hinz TK, Kleczko E, Singleton KR, Marek LA, Helfrich BA, Cummings CT, Graham DK, Astling D, Tan AC and Heasley LE: A mechanism of resistance to gefitinib mediated by cellular reprogramming and the acquisition of an FGF2-FGFR1 autocrine growth loop. Oncogenesis. 2:e392013. View Article : Google Scholar : PubMed/NCBI
17	Yao Z, Fenoglio S, Gao DC, Camiolo M, Stiles B, Lindsted T, Schlederer M, Johns C, Altorki N, Mittal V, et al: TGF-beta IL-6 axis mediates selective and adaptive mechanisms of resistance to molecular targeted therapy in lung cancer. Proc Natl Acad Sci USA. 107:15535–15540. 2010. View Article : Google Scholar : PubMed/NCBI
18	Yan HQ, Huang XB, Ke SZ, Jiang YN, Zhang YH, Wang YN, Li J and Gao FG: Interleukin 6 augments lung cancer chemotherapeutic resistance via ataxia-telangiectasia mutated/NF-kappaB pathway activation. Cancer Sci. 105:1220–1227. 2014. View Article : Google Scholar : PubMed/NCBI
19	Liu CC, Lin JH, Hsu TW, Su K, Li AF, Hsu HS and Hung SC: IL-6 enriched lung cancer stem-like cell population by inhibition of cell cycle regulators via DNMT1 upregulation. Int J Cancer. 136:547–559. 2015.PubMed/NCBI
20	Yi H, Cho HJ, Cho SM, Jo K, Park JA, Kim NH, Amidon GL, Kim JS and Shin HC: Blockade of interleukin-6 receptor suppresses the proliferation of H460 lung cancer stem cells. Int J Oncol. 41:310–316. 2012.PubMed/NCBI
21	Templeton AK, Miyamoto S, Babu A, Munshi A and Ramesh R: Cancer stem cells: Progress and challenges in lung cancer. Stem Cell Investig. 1:92014.PubMed/NCBI
22	Godin-Heymann N, Ulkus L, Brannigan BW, McDermott U, Lamb J, Maheswaran S, Settleman J and Haber DA: The T790M ‘gatekeeper’ mutation in EGFR mediates resistance to low concentrations of an irreversible EGFR inhibitor. Mol Cancer Ther. 7:874–879. 2008. View Article : Google Scholar : PubMed/NCBI
23	Engelman JA, Zejnullahu K, Mitsudomi T, Song Y, Hyland C, Park JO, Lindeman N, Gale CM, Zhao X, Christensen J, et al: MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. Science. 316:1039–1043. 2007. View Article : Google Scholar : PubMed/NCBI
24	Rosell R, Karachaliou N, Morales-Espinosa D, Costa C, Molina MA, Sansano I, Gasco A, Viteri S, Massuti B, Wei J, et al: Adaptive resistance to targeted therapies in cancer. Transl Lung Cancer Res. 2:152–159. 2013.PubMed/NCBI
25	Li L, Han R, Xiao H, Lin C, Wang Y, Liu H, Li K, Chen H, Sun F, Yang Z, et al: Metformin sensitizes EGFR-TKI-resistant human lung cancer cells in vitro and in vivo through inhibition of IL-6 signaling and EMT reversal. Clin Cancer Res. 20:2714–2726. 2014. View Article : Google Scholar : PubMed/NCBI
26	Kumar M, Allison DF, Baranova NN, Wamsley JJ, Katz AJ, Bekiranov S, Jones DR and Mayo MW: NF-κB regulates mesenchymal transition for the induction of non-small cell lung cancer initiating cells. PLoS One. 8:e685972013. View Article : Google Scholar : PubMed/NCBI
27	Min C, Eddy SF, Sherr DH and Sonenshein GE: NF-kappaB and epithelial to mesenchymal transition of cancer. J Cell Biochem. 104:733–744. 2008. View Article : Google Scholar : PubMed/NCBI
28	Akunuru S, James Zhai Q and Zheng Y: Non-small cell lung cancer stem/progenitor cells are enriched in multiple distinct phenotypic subpopulations and exhibit plasticity. Cell Death Dis. 3:e3522012. View Article : Google Scholar : PubMed/NCBI
29	Chang TS, Chen CL, Wu YC, Liu JJ, Kuo YC, Lee KF, Lin SY, Lin SE, Tung SY, Kuo LM, et al: Inflammation promotes expression of stemness-related properties in HBV-related hepatocellular carcinoma. PLoS One. 11:e01498972016. View Article : Google Scholar : PubMed/NCBI
30	Medema JP: Cancer stem cells: The challenges ahead. Nat Cell Biol. 15:338–344. 2013. View Article : Google Scholar : PubMed/NCBI
31	Lin L, Hutzen B, Lee HF, Peng Z, Wang W, Zhao C, Lin HJ, Sun D, Li PK, Li C, et al: Evaluation of STAT3 signaling in ALDH+ and ALDH+/CD44+/CD24− subpopulations of breast cancer cells. PLoS One. 8:e828212013. View Article : Google Scholar : PubMed/NCBI
32	Shao C, Sullivan JP, Girard L, Augustyn A, Yenerall P, Rodriguez-Canales J, Liu H, Behrens C, Shay JW, Wistuba II and Minna JD: Essential role of aldehyde dehydrogenase 1A3 for the maintenance of non-small cell lung cancer stem cells is associated with the STAT3 pathway. Clin Cancer Res. 20:4154–4166. 2014. View Article : Google Scholar : PubMed/NCBI
33	van der Zee M, Sacchetti A, Cansoy M, Joosten R, Teeuwssen M, Heijmans-Antonissen C, Ewing-Graham PC, Burger CW, Blok LJ and Fodde R: IL6/JAK1/STAT3 signaling blockade in endometrial cancer affects the ALDHhi/CD126+ stem-like component and reduces tumor burden. Cancer Res. 75:3608–3622. 2015. View Article : Google Scholar : PubMed/NCBI
34	Han Z, Wang X, Ma L, Chen L, Xiao M, Huang L, Cao Y, Bai J, Ma D, Zhou J and Hong Z: Inhibition of STAT3 signaling targets both tumor-initiating and differentiated cell populations in prostate cancer. Oncotarget. 5:8416–8428. 2014. View Article : Google Scholar : PubMed/NCBI
35	Lin L, Fuchs J, Li C, Olson V, Bekaii-Saab T and Lin J: STAT3 signaling pathway is necessary for cell survival and tumorsphere forming capacity in ALDH⁺/CD133⁺ stem cell-like human colon cancer cells. Biochem Biophys Res Commun. 416:246–251. 2011. View Article : Google Scholar : PubMed/NCBI
36	Chang CH, Hsiao CF, Yeh YM, Chang GC, Tsai YH, Chen YM, Huang MS, Chen HL, Li YJ, Yang PC, et al: Circulating interleukin-6 level is a prognostic marker for survival in advanced nonsmall cell lung cancer patients treated with chemotherapy. Int J Cancer. 132:1977–1985. 2013. View Article : Google Scholar : PubMed/NCBI
37	Hennigan S and Kavanaugh A: Interleukin-6 inhibitors in the treatment of rheumatoid arthritis. Ther Clin Risk Manag. 4:767–775. 2008. View Article : Google Scholar : PubMed/NCBI
38	Sebba A: Tocilizumab: The first interleukin-6-receptor inhibitor. Am J Health Syst Pharm. 65:1413–1418. 2008. View Article : Google Scholar : PubMed/NCBI
39	Bayliss TJ, Smith JT, Schuster M, Dragnev KH and Rigas JR: A humanized anti-IL-6 antibody (ALD518) in non-small cell lung cancer. Expert Opin Biol Ther. 11:1663–1668. 2011. View Article : Google Scholar : PubMed/NCBI