Abstract
Purpose
To study an association between IL-6 signaling and resistance to radiotherapy of prostate cancer (PCa) and explore the molecular mechanisms involved.
Methods
IL-6 expressing C4-2 and CWR22Rv1 (C4-2IL-6/CWRIL-6) and vector control (C4-2vec/CWRvec) cell lines were developed. Radiation-sensitivities of these cells were compared in clonogenic assay, Comet assay, and γH2AX staining. In xenograft animal studies, radiation-sensitivity of C4-2IL-6 cell-derived tumors vs. C4-2vec cell-derived tumors was investigated. To reveal IL-6 downstream molecules involved in DNA repair after radiation, qPCR and Western blot analyses as well as immunofluorescence staining were performed. Transcriptional control of IL-6 on ATM and ATR molecules was also investigated.
Results
We found C4-2IL-6 and CWRIL-6 cells survived better than their vector control cells after irradiation, and animal studies confirmed such in vitro results. We discovered that DNA repair-related molecules such as ATM, ATR, BRCA1, and BRCA2 were significantly upregulated in irradiated IL-6 expressing cells compared with vector control cells. We further defined that IL-6 signaling regulated cellular expressions of ATM and ATR at the transcriptional level through the activation of Stat3 signaling pathway.
Conclusions
IL-6 leads to PCa resistance to radiation through upregulation of DNA repair associated molecules ATM, ATR, BRCA1, and BRCA2.
Similar content being viewed by others
References
Bunz F (2016) Principles of cancer genetics. Anticancer Res 36:4979
Bellucci R, Martin A, Bommarito D, Wang K, Hansen SH, Freeman GJ, Ritz J (2015) Interferon-gamma-induced activation of JAK1 and JAK2 suppresses tumor cell susceptibility to NK cells through upregulation of PD-L1 expression. Oncoimmunology 4:e1008824. https://doi.org/10.1080/2162402x.2015.1008824
Bonkhoff H (2012) Factors implicated in radiation therapy failure and radiosensitization of prostate cancer. Prostate Cancer 2012:593241. https://doi.org/10.1155/2012/593241
Bonner WM, Redon CE, Dickey JS, Nakamura AJ, Sedelnikova OA, Solier S, Pommier Y (2008) GammaH2AX and cancer. Nat Rev Cancer 8:957–967. https://doi.org/10.1038/nrc2523
Cazzalini O, Scovassi AI, Savio M, Stivala LA, Prosperi E (2010) Multiple roles of the cell cycle inhibitor p21(CDKN1A) in the DNA damage response. Mutat Res 704:12–20. https://doi.org/10.1016/j.mrrev.2010.01.009
Chang PC et al (2014) Autophagy pathway is required for IL-6 induced neuroendocrine differentiation and chemoresistance of prostate cancer LNCaP cells. PLoS One 9:e88556. https://doi.org/10.1371/journal.pone.0088556
Chen MF, Hsieh CC, Chen WC, Lai CH (2012) Role of interleukin-6 in the radiation response of liver tumors. Int J Radiat Oncol Biol Phys 84:e621–e630. https://doi.org/10.1016/j.ijrobp.2012.07.2360
Chen N et al (2015a) Upregulation of PD-L1 by EGFR activation mediates the immune escape in EGFR-Driven NSCLC: implication for optional immune targeted therapy for NSCLC patients with EGFR mutation. J Thorac Oncol 10:910–923. https://doi.org/10.1097/jto.0000000000000500
Chen Y et al (2015b) IL-6 signaling promotes DNA repair and prevents apoptosis in CD133+ stem-like cells of lung cancer after radiation. Radiat Oncol 10:227. https://doi.org/10.1186/s13014-015-0534-1
Cheng A et al (2018) ATM and ATR play complementary roles in the behavior of excitatory and inhibitory vesicle populations. Proc Natl Acad Sci USA 115:E292–E301. https://doi.org/10.1073/pnas.1716892115
Culig Z, Puhr M (2012) Interleukin-6: a multifunctional targetable cytokine in human prostate cancer. Mol Cell Endocrinol 360:52–58. https://doi.org/10.1016/j.mce.2011.05.033
Culig Z, Bartsch G, Hobisch A (2002) Interleukin-6 regulates androgen receptor activity and prostate cancer cell growth. Mol Cell Endocrinol 197:231–238
Feng Z, Kachnic L, Zhang J, Powell SN, Xia F (2004) DNA damage induces p53-dependent BRCA1 nuclear export. J Biol Chem 279:28574–28584. https://doi.org/10.1074/jbc.m404137200
Figueroa-Gonzalez G, Perez-Plasencia C (2017) Strategies for the evaluation of DNA damage and repair mechanisms in cancer. Oncol Lett 13:3982–3988. https://doi.org/10.3892/ol.2017.6002
Flynn RL, Zou L (2011) ATR: a master conductor of cellular responses to DNA replication stress. Trends Biochem Sci 36:133–140. https://doi.org/10.1016/j.tibs.2010.09.005
Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C (2006) Clonogenic assay of cells in vitro. Nat Protoc 1:2315–2319. https://doi.org/10.1038/nprot.2006.339
Fujita Y et al (2015) The clinical relevance of the miR-197/CKS1B/STAT3-mediated PD-L1 network in chemoresistant non-small-cell lung cancer. Mol Ther 23:717–727. https://doi.org/10.1038/mt.2015.10
Giri D, Ozen M, Ittmann M (2001) Interleukin-6 is an autocrine growth factor in human prostate cancer. Am J Pathol 159:2159–2165. https://doi.org/10.1016/s0002-9440(10)63067-2
Gowrishankar K, Gunatilake D, Gallagher SJ, Tiffen J, Rizos H, Hersey P (2015) Inducible but not constitutive expression of PD-L1 in human melanoma cells is dependent on activation of NF-kappaB. PloS One 10:e0123410. https://doi.org/10.1371/journal.pone.0123410
Hayden AJ, Catton C, Pickles T (2010) Radiation therapy in prostate cancer: a risk-adapted strategy. Curr Oncol 17(Suppl 2):S18–S24
Henderson BR (2005) Regulation of BRCA1, BRCA2 and BARD1 intracellular trafficking. Bioessays 27:884–893. https://doi.org/10.1002/bies.20277
Ikeda S et al (2016) PD-L1 Is Upregulated by simultaneous amplification of the PD-L1 and JAK2 genes in non-small cell lung cancer. J Thorac Oncol 11:62–71. https://doi.org/10.1016/j.jtho.2015.09.010
Ji J et al (2017) Phosphorylated fraction of H2AX as a measurement for DNA damage in cancer cells and potential applications of a novel assay. PloS One 12:e0171582. https://doi.org/10.1371/journal.pone.0171582
Johnson DE, O’Keefe RA, Grandis JR (2018) Targeting the IL-6/JAK/STAT3 signalling axis in cancer. Nat Rev Clin Oncol 15:234–248. https://doi.org/10.1038/nrclinonc.2018.8
Kidiyoor GR, Kumar A, Foiani M (2016) ATR-mediated regulation of nuclear and cellular plasticity. DNA Repair (Amst) 44:143–150. https://doi.org/10.1016/j.dnarep.2016.05.020
Lastwika KJ et al (2016) Control of PD-L1 expression by oncogenic activation of the AKT–mTOR pathway in non-small cell lung cancer. Cancer Res 76:227–238. https://doi.org/10.1158/0008-5472.can-14-3362
Leisching G, Loos B, Botha M, Engelbrecht AM (2015) Bcl-2 confers survival in cisplatin treated cervical cancer cells: circumventing cisplatin dose-dependent toxicity and resistance. J Transl Med 13:328. https://doi.org/10.1186/s12967-015-0689-4
Lin DL, Whitney MC, Yao Z, Keller ET (2001) Interleukin-6 induces responsiveness in prostate cancer cells through up-regulation of androgen receptor expression. Clin Cancer Res 7:1773–1781
Lohse I et al (2015) BRCA1 and BRCA2 mutations sensitize to chemotherapy in patient-derived pancreatic cancer xenografts. Br J Cancer 113:425–432. https://doi.org/10.1038/bjc.2015.220
Manic G, Obrist F, Sistigu A, Vitale I (2015) Trial watch: targeting ATM–CHK2 and ATR–CHK1 pathways for anticancer therapy. Mol Cell Oncol 2:e1012976. https://doi.org/10.1080/23723556.2015.1012976
Marzec M et al (2008) Oncogenic kinase NPM/ALK induces through STAT3 expression of immunosuppressive protein CD274 (PD-L1, B7-H1). Proc Natl Acad Sci USA 105:20852–20857. https://doi.org/10.1073/pnas.0810958105
Matsuoka Y et al (2016) IL-6 controls resistance to radiation by suppressing oxidative stress via the Nrf2-antioxidant pathway in oral squamous cell carcinoma. Br J Cancer 115:1234–1244. https://doi.org/10.1038/bjc.2016.327
Nguyen DP, Li J, Tewari AK (2014) Inflammation and prostate cancer: the role of interleukin 6 (IL-6). BJU Int 113:986–992. https://doi.org/10.1111/bju.12452
Noh H, Hu J, Wang X, Xia X, Satelli A, Li S (2015) Immune checkpoint regulator PD-L1 expression on tumor cells by contacting CD11b positive bone marrow derived stromal cells. Cell Commun Signal 13:14. https://doi.org/10.1186/s12964-015-0093-y
Oh DY et al (2015) Phase I study of OPB-31121, an oral STAT3 Inhibitor, in patients with advanced solid tumors. Cancer Res Treat 47:607–615. https://doi.org/10.4143/crt.2014.249
Sanford DC, DeWille JW (2005) C/EBPdelta is a downstream mediator of IL-6 induced growth inhibition of prostate cancer cells. Prostate 63:143–154. https://doi.org/10.1002/pros.20159
Scheller J, Chalaris A, Schmidt-Arras D, Rose-John S (2011) The pro- and anti-inflammatory properties of the cytokine interleukin-6. Biochimica et Biophysica Acta 1813:878–888. https://doi.org/10.1016/j.bbamcr.2011.01.034
Shiloh Y, Ziv Y (2013) The ATM protein kinase: regulating the cellular response to genotoxic stress, and more. Nat Rev Mol Cell Biol 14:197–210. https://doi.org/10.1038/nrm3546
Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191
Smith J, Tho LM, Xu N, Gillespie DA (2010) The ATM–Chk2 and ATR–Chk1 pathways in DNA damage signaling and cancer. Adv Cancer Res 108:73–112. https://doi.org/10.1016/b978-0-12-380888-2.00003-0
Tamari Y, Kashino G, Mori H (2017) Acquisition of radioresistance by IL-6 treatment is caused by suppression of oxidative stress derived from mitochondria after gamma-irradiation. J Radiat Res 58:412–420. https://doi.org/10.1093/jrr/rrw084
Vanneste BG, Van Limbergen EJ, van Lin EN, van Roermund JG, Lambin P (2016) Prostate cancer radiation therapy: what do clinicians have to know? Biomed Res Int 2016:6829875. https://doi.org/10.1155/2016/6829875
Vequaud E, Desplanques G, Jezequel P, Juin P, Barille-Nion S (2016) Survivin contributes to DNA repair by homologous recombination in breast cancer cells. Breast Cancer Res Treat 155:53–63. https://doi.org/10.1007/s10549-015-3657-z
Volcic M, Karl S, Baumann B, Salles D, Daniel P, Fulda S, Wiesmuller L (2012) NF-kappaB regulates DNA double-strand break repair in conjunction with BRCA1-CtIP complexes. Nucleic Acids Res 40:181–195. https://doi.org/10.1093/nar/gkr687
Weber AM, Ryan AJ (2015) ATM and ATR as therapeutic targets in cancer. Pharmacol Ther 149:124–138. https://doi.org/10.1016/j.pharmthera.2014.12.001
Williams AB, Schumacher B (2016) p53 in the DNA-damage-repair process. Cold Spring Harb Perspect Med. https://doi.org/10.1101/cshperspect.a026070
Wu CT, Chen MF, Chen WC, Hsieh CC (2013) The role of IL-6 in the radiation response of prostate cancer. Radiat Oncol 8:159. https://doi.org/10.1186/1748-717x-8-159
Xu C et al (2014) Loss of Lkb1 and Pten leads to lung squamous cell carcinoma with elevated PD-L1 expression. Cancer Cell 25:590–604. https://doi.org/10.1016/j.ccr.2014.03.033
Yamamoto R et al (2009) B7-H1 expression is regulated by MEK/ERK signaling pathway in anaplastic large cell lymphoma and Hodgkin lymphoma. Cancer Sci 100:2093–2100. https://doi.org/10.1111/j.1349-7006.2009.01302.x
Yano K, Morotomi K, Saito H, Kato M, Matsuo F, Miki Y (2000) Nuclear localization signals of the BRCA2 protein. Biochem Biophys Res Commun 270:171–175. https://doi.org/10.1006/bbrc.2000.2392
Yoon JH, Ahn SG, Lee BH, Jung SH, Oh SH (2012) Role of autophagy in chemoresistance: regulation of the ATM-mediated DNA-damage signaling pathway through activation of DNA-PKcs and PARP-1. Biochem Pharmacol 83:747–757. https://doi.org/10.1016/j.bcp.2011.12.029
Zang C et al (2017) IL-6/STAT3/TWIST inhibition reverses ionizing radiation-induced EMT and radioresistance in esophageal squamous carcinoma. Oncotarget 8:11228–11238. https://doi.org/10.18632/oncotarget.14495
Zhao R, Yang FT, Alexander DR (2004) An oncogenic tyrosine kinase inhibits DNA repair and DNA-damage-induced Bcl-xL deamidation in T cell transformation. Cancer Cell 5:37–49
Zheng X-HL, Wang N, Zhou H, Ma W-J, Zhang T-C (2015) Construction and functional analysis of luciferase reporter plasmids containing ATM and ATR gene promoters. Adv Appl Biotechnol Lect Notes Electr Eng Chapter 65:627–634
Acknowledgements
We thank Mrs. Laura Finger for the help with manuscript preparation. This work was supported by the Richard T. Bell Endowed Professorship (YC).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no conflicts of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chen, X., Chen, F., Ren, Y. et al. IL-6 signaling contributes to radioresistance of prostate cancer through key DNA repair-associated molecules ATM, ATR, and BRCA 1/2. J Cancer Res Clin Oncol 145, 1471–1484 (2019). https://doi.org/10.1007/s00432-019-02917-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-019-02917-z