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Breast Cancer Research
Published in: Breast Cancer Research 2/2011

Open Access 01-04-2011 | Research article

Impact of oncogenic K-RASon YB-1 phosphorylation induced by ionizing radiation

Authors: Mahmoud Toulany, Tim-Andre Schickfluß, Wolfgang Eicheler, Rainer Kehlbach, Birgit Schittek, H Peter Rodemann

Published in: Breast Cancer Research | Issue 2/2011

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Abstract

Introduction

Expression of Y-box binding protein-1 (YB-1) is associated with tumor progression and drug resistance. Phosphorylation of YB-1 at serine residue 102 (S102) in response to growth factors is required for its transcriptional activity and is thought to be regulated by cytoplasmic signaling phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathways. These pathways can be activated by growth factors and by exposure to ionizing radiation (IR). So far, however, no studies have been conducted on IR-induced YB-1 phosphorylation.

Methods

IR-induced YB-1 phosphorylation in K-RAS wild-type (K-RASwt) and K-RAS-mutated (K-RASmt) breast cancer cell lines was investigated. Using pharmacological inhibitors, small interfering RNA (siRNA) and plasmid-based overexpression approaches, we analyzed pathways involved in YB-1 phosphorylation by IR. Using γ-H2AX foci and standard colony formation assays, we investigated the function of YB-1 in repair of IR-induced DNA double-stranded breaks (DNA-DSB) and postirradiation survival was investigated.

Results

The average level of phosphorylation of YB-1 in the breast cancer cell lines SKBr3, MCF-7, HBL100 and MDA-MB-231 was significantly higher than that in normal cells. Exposure to IR and stimulation with erbB1 ligands resulted in phosphorylation of YB-1 in K-RASwt SKBr3, MCF-7 and HBL100 cells, which was shown to be K-Ras-independent. In contrast, lack of YB-1 phosphorylation after stimulation with either IR or erbB1 ligands was observed in K-RASmt MDA-MB-231 cells. Similarly to MDA-MB-231 cells, YB-1 became constitutively phosphorylated in K-RASwt cells following the overexpression of mutated K-RAS, and its phosphorylation was not further enhanced by IR. Phosphorylation of YB-1 as a result of irradiation or K-RAS mutation was dependent on erbB1 and its downstream pathways, PI3K and MAPK/ERK. In K-RASmt cells K-RAS siRNA as well as YB-1 siRNA blocked repair of DNA-DSB. Likewise, YB-1 siRNA increased radiation sensitivity.

Conclusions

IR induces YB-1 phosphorylation. YB-1 phosphorylation induced by oncogenic K-Ras or IR enhances repair of DNA-DSB and postirradiation survival via erbB1 downstream PI3K/Akt and MAPK/ERK signaling pathways.
Appendix
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Literature
1.
Bargou RC, Jürchott K, Wagener C, Bergmann S, Metzner S, Bommert K, Mapara MY, Winzer KJ, Dietel M, Dörken B, Royer HD: Nuclear localization and increased levels of transcription factor YB-1 in primary human breast cancers are associated with intrinsic MDR1 gene expression. Nat Med. 1997, 3: 447-450. 10.1038/nm0497-447.PubMed
2.
Yu YN, Yip GW, Tan PH, Thike AA, Matsumoto K, Tsujimoto M, Bay BH: Y-box binding protein 1 is up-regulated in proliferative breast cancer and its inhibition deregulates the cell cycle. Int J Oncol. 2010, 37: 483-492.PubMed
3.
To K, Fotovati A, Reipas KM, Law JH, Hu K, Wang J, Astanehe A, Davies AH, Lee L, Stratford AL, Raouf A, Johnson P, Berquin IM, Royer HD, Eaves CJ, Dunn SE: Y-box binding protein-1 induces the expression of CD44 and CD49f leading to enhanced self-renewal, mammosphere growth, and drug resistance. Cancer Res. 2010, 70: 2840-2851. 10.1158/0008-5472.CAN-09-3155.PubMedPubMedCentral
4.
Yang JY, Ha SA, Yang YS, Kim JW: p-Glycoprotein ABCB5 and YB-1 expression plays a role in increased heterogeneity of breast cancer cells: correlations with cell fusion and doxorubicin resistance. BMC Cancer. 2010, 10: 388-10.1186/1471-2407-10-388.PubMedPubMedCentral
5.
Janz M, Harbeck N, Dettmar P, Berger U, Schmidt A, Jürchott K, Schmitt M, Royer HD: Y-box factor YB-1 predicts drug resistance and patient outcome in breast cancer independent of clinically relevant tumor biologic factors HER2, uPA and PAI-1. Int J Cancer. 2002, 97: 278-282. 10.1002/ijc.1610.PubMed
6.
Basaki Y, Taguchi K, Izumi H, Murakami Y, Kubo T, Hosoi F, Watari K, Nakano K, Kawaguchi H, Ohno S, Kohno K, Ono M, Kuwano M: Y-box binding protein-1 (YB-1) promotes cell cycle progression through CDC6-dependent pathway in human cancer cells. Eur J Cancer. 2010, 46: 954-965. 10.1016/j.ejca.2009.12.024.PubMed
7.
Stratford AL, Habibi G, Astanehe A, Jiang H, Hu K, Park E, Shadeo A, Buys TP, Lam W, Pugh T, Marra M, Nielsen TO, Klinge U, Mertens PR, Aparicio S, Dunn SE: 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. 2007, 9: R61-10.1186/bcr1767.PubMedPubMedCentral
8.
Fujii T, Kawahara A, Basaki Y, Hattori S, Nakashima K, Nakano K, Shirouzu K, Kohno K, Yanagawa T, Yamana H, Nishio K, Ono M, Kuwano M, Kage M: Expression of HER2 and estrogen receptor α depends upon nuclear localization of Y-box binding protein-1 in human breast cancers. Cancer Res. 2008, 68: 1504-1512. 10.1158/0008-5472.CAN-07-2362.PubMed
9.
Lee C, Dhillon J, Wang MY, Gao Y, Hu K, Park E, Astanehe A, Hung MC, Eirew P, Eaves CJ, Dunn SE: Targeting YB-1 in HER-2 overexpressing breast cancer cells induces apoptosis via the mTOR/STAT3 pathway and suppresses tumor growth in mice. Cancer Res. 2008, 68: 8661-8666. 10.1158/0008-5472.CAN-08-1082.PubMed
10.
Kashihara M, Azuma K, Kawahara A, Basaki Y, Hattori S, Yanagawa T, Terazaki Y, Takamori S, Shirouzu K, Aizawa H, Nakano K, Kage M, Kuwano M, Ono M: Nuclear Y-box binding protein-1, a predictive marker of prognosis, is correlated with expression of HER2/ErbB2 and HER3/ErbB3 in non-small cell lung cancer. J Thorac Oncol. 2009, 4: 1066-1074. 10.1097/JTO.0b013e3181ae2828.PubMed
11.
Stratford AL, Fry CJ, Desilets C, Davies AH, Cho YY, Li Y, Dong Z, Berquin IM, Roux PP, Dunn SE: Y-box binding protein-1 serine 102 is a downstream target of p90 ribosomal S6 kinase in basal-like breast cancer cells. Breast Cancer Res. 2008, 10: R99-10.1186/bcr2202.PubMedPubMedCentral
12.
Sutherland BW, Kucab J, Wu J, Lee C, Cheang MC, Yorida E, Turbin D, Dedhar S, Nelson C, Pollak M, Leighton Grimes H, Miller K, Badve S, Huntsman D, Blake-Gilks C, Chen M, Pallen CJ, Dunn SE: Akt phosphorylates the Y-box binding protein 1 at Ser102 located in the cold shock domain and affects the anchorage-independent growth of breast cancer cells. Oncogene. 2005, 24: 4281-4292. 10.1038/sj.onc.1208590.PubMed
13.
Law JH, Li Y, To K, Wang M, Astanehe A, Lambie K, Dhillon J, Jones SJ, Gleave ME, Eaves CJ, Dunn SE: Molecular decoy to the Y-box binding protein-1 suppresses the growth of breast and prostate cancer cells whilst sparing normal cell viability. PLoS One. 2010, 5: 10.1371/journal.pone.0012661. pii:e12661
14.
Kalra J, Sutherland BW, Stratford AL, Dragowska W, Gelmon KA, Dedhar S, Dunn SE, Bally MB: Suppression of Her2/neu expression through ILK inhibition is regulated by a pathway involving TWIST and YB-1. Oncogene. 2010, 29: 6343-6356. 10.1038/onc.2010.366.PubMedPubMedCentral
15.
Mukherjee B, McEllin B, Camacho CV, Tomimatsu N, Sirasanagandala S, Nannepaga S, Hatanpaa KJ, Mickey B, Madden C, Maher E, Boothman DA, Furnari F, Cavenee WK, Bachoo RM, Burma S: EGFRvIII and DNA double-strand break repair: a molecular mechanism for radioresistance in glioblastoma. Cancer Res. 2009, 69: 4252-4259. 10.1158/0008-5472.CAN-08-4853.PubMedPubMedCentral
16.
Milas L, Fan Z, Andratschke NH, Ang KK: Epidermal growth factor receptor and tumor response to radiation: in vivo preclinical studies. Int J Radiat Oncol Biol Phys. 2004, 58: 966-971. 10.1016/j.ijrobp.2003.08.035.PubMed
17.
Sartor CI: Biological modifiers as potential radiosensitizers: targeting the epidermal growth factor receptor family. Semin Oncol. 2000, 27 (6 Suppl 11): 15-20. discussion 92-100PubMed
18.
Nyati MK, Morgan MA, Feng FY, Lawrence TS: Integration of EGFR inhibitors with radiochemotherapy. Nat Rev Cancer. 2006, 6: 876-885. 10.1038/nrc1953.PubMed
19.
Baumann M, Krause M, Dikomey E, Dittmann K, Dörr W, Kasten-Pisula U, Rodemann HP: EGFR-targeted anti-cancer drugs in radiotherapy: preclinical evaluation of mechanisms. Radiother Oncol. 2007, 83: 238-248. 10.1016/j.radonc.2007.04.006.PubMed
20.
Harari PM: Epidermal growth factor receptor inhibition strategies in oncology. Endocr Relat Cancer. 2004, 11: 689-708. 10.1677/erc.1.00600.PubMed
21.
Harari PM, Huang S: Radiation combined with EGFR signal inhibitors: head and neck cancer focus. Semin Radiat Oncol. 2006, 16: 38-44. 10.1016/j.semradonc.2005.08.005.PubMed
22.
Bonner JA, Harari PM, Giralt J, Azarnia N, Shin DM, Cohen RB, Jones CU, Sur R, Raben D, Jassem J, Ove R, Kies MS, Baselga J, Youssoufian H, Amellal N, Rowinsky EK, Ang KK: Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. N Engl J Med. 2006, 354: 567-578. 10.1056/NEJMoa053422.PubMed
23.
Shepard HM, Brdlik CM, Schreiber H: Signal integration: a framework for understanding the efficacy of therapeutics targeting the human EGFR family. J Clin Invest. 2008, 118: 3574-3581. 10.1172/JCI36049.PubMedPubMedCentral
24.
Lee HC, An S, Lee H, Woo SH, Jin HO, Seo SK, Choe TB, Yoo DH, Lee SJ, Hong YJ, Park MJ, Rhee CH, Park IC, Hong SI: Activation of epidermal growth factor receptor and its downstream signaling pathway by nitric oxide in response to ionizing radiation. Mol Cancer Res. 2008, 6: 996-1002. 10.1158/1541-7786.MCR-08-0113.PubMed
25.
Toulany M, Dittmann K, Baumann M, Rodemann HP: Radiosensitization of Ras-mutated human tumor cells in vitro by the specific EGF receptor antagonist BIBX1382BS. Radiother Oncol. 2005, 74: 117-129. 10.1016/j.radonc.2004.11.008.PubMed
26.
Toulany M, Rodemann HP: Membrane receptor signaling and control of DNA repair after exposure to ionizing radiation. Nuklearmedizin. 2010, 49 (Suppl 1): S26-S30.PubMed
27.
Rodemann HP, Dittmann K, Toulany M: Radiation-induced EGFR-signaling and control of DNA-damage repair. Int J Radiat Biol. 2007, 83: 781-791. 10.1080/09553000701769970.PubMed
28.
Hynes NE, MacDonald G: ErbB receptors and signaling pathways in cancer. Curr Opin Cell Biol. 2009, 21: 177-184. 10.1016/j.ceb.2008.12.010.PubMed
29.
Cengel KA, Voong KR, Chandrasekaran S, Maggiorella L, Brunner TB, Stanbridge E, Kao GD, McKenna WG, Bernhard EJ: Oncogenic K-Ras signals through epidermal growth factor receptor and wild-type H-Ras to promote radiation survival in pancreatic and colorectal carcinoma cells. Neoplasia. 2007, 9: 341-348. 10.1593/neo.06823.PubMedPubMedCentral
30.
Toulany M, Baumann M, Rodemann HP: Stimulated PI3K-AKT signaling mediated through ligand or radiation-induced EGFR depends indirectly, but not directly, on constitutive K-Ras activity. Mol Cancer Res. 2007, 5: 863-872. 10.1158/1541-7786.MCR-06-0297.PubMed
31.
Dreissigacker U, Mueller MS, Unger M, Siegert P, Genze F, Gierschik P, Giehl K: Oncogenic K-Ras down-regulates Rac1 and RhoA activity and enhances migration and invasion of pancreatic carcinoma cells through activation of p38. Cell Signal. 2006, 18: 1156-1168. 10.1016/j.cellsig.2005.09.004.PubMed
32.
Toulany M, Minjgee M, Kehlbach R, Chen J, Baumann M, Rodemann HP: ErbB2 expression through heterodimerization with erbB1 is necessary for ionizing radiation but not EGF-induced activation of Akt survival pathway. Radiother Oncol. 2010, 97: 338-345. 10.1016/j.radonc.2010.03.008.PubMed
33.
Toulany M, Dittmann K, Fehrenbacher B, Schaller M, Baumann M, Rodemann HP: PI3K-Akt signaling regulates basal, but MAP-kinase signaling regulates radiation-induced XRCC1 expression in human tumor cells in vitro. DNA Repair (Amst). 2008, 7: 1746-1756. 10.1016/j.dnarep.2008.06.017.
34.
Toulany M, Kasten-Pisula U, Brammer I, Wang S, Chen J, Dittmann K, Baumann M, Dikomey E, Rodemann HP: Blockage of epidermal growth factor receptor-phosphatidylinositol 3-kinase-AKT signaling increases radiosensitivity of K-RAS mutated human tumor cells in vitro by affecting DNA repair. Clin Cancer Res. 2006, 12: 4119-4126. 10.1158/1078-0432.CCR-05-2454.PubMed
35.
Evdokimova V, Ruzanov P, Anglesio MS, Sorokin AV, Ovchinnikov LP, Buckley J, Triche TJ, Sonenberg N, Sorensen PH: Akt-mediated YB-1 phosphorylation activates translation of silent mRNA species. Mol Cell Biol. 2006, 26: 277-292. 10.1128/MCB.26.1.277-292.2006.PubMedPubMedCentral
36.
Contessa JN, Hampton J, Lammering G, Mikkelsen RB, Dent P, Valerie K, Schmidt-Ullrich RK: Ionizing radiation activates Erb-B receptor dependent Akt and p70 S6 kinase signaling in carcinoma cells. Oncogene. 2002, 21: 4032-4041. 10.1038/sj.onc.1205500.PubMed
37.
Eckert LB, Repasky GA, Ülkü AS, McFall A, Zhou H, Sartor CI, Der CJ: Involvement of Ras activation in human breast cancer cell signaling, invasion, and anoikis. Cancer Res. 2004, 64: 4585-4592. 10.1158/0008-5472.CAN-04-0396.PubMed
38.
Egeblad M, Mortensen OH, van Kempen LC, Jäättelä M: BIBX1382BS, but not AG1478 or PD153035 inhibits the ErbB kinases at different concentrations in intact cells. Biochem Biophys Res Commun. 2001, 281: 25-31. 10.1006/bbrc.2001.4302.PubMed
39.
Gaudreault I, Guay D, Lebel M: YB-1 promotes strand separation in vitro of duplex DNA containing either mispaired bases or cisplatin modifications, exhibits endonucleolytic activities and binds several DNA repair proteins. Nucleic Acids Res. 2004, 32: 316-327. 10.1093/nar/gkh170.PubMedPubMedCentral
40.
Hasegawa SL, Doetsch PW, Hamilton KK, Martin AM, Okenquist SA, Lenz J, Boss JM: DNA binding properties of YB-1 and dbpA: binding to double-stranded, single-stranded, and abasic site containing DNAs. Nucleic Acids Res. 1991, 19: 4915-4920. 10.1093/nar/19.18.4915.PubMedPubMedCentral
41.
Fuhrman CB, Kilgore J, LaCoursiere YD, Lee CM, Milash BA, Soisson AP, Zempolich KA: Radiosensitization of cervical cancer cells via double-strand DNA break repair inhibition. Gynecol Oncol. 2008, 110: 93-98. 10.1016/j.ygyno.2007.08.073.PubMed
42.
Choi EJ, Ryu YK, Kim SY, Wu HG, Kim JS, Kim IH, Kim IA: Targeting epidermal growth factor receptor-associated signaling pathways in non-small cell lung cancer cells: implication in radiation response. Mol Cancer Res. 2010, 8: 1027-1036. 10.1158/1541-7786.MCR-09-0507.PubMed
43.
Hamada N, Matsumoto H, Hara T, Kobayashi Y: Intercellular and intracellular signaling pathways mediating ionizing radiation-induced bystander effects. J Radiat Res (Tokyo). 2007, 48: 87-95. 10.1269/jrr.06084.
44.
Wu J, Lee C, Yokom D, Jiang H, Cheang MC, Yorida E, Turbin D, Berquin IM, Mertens PR, Iftner T, Gilks CB, Dunn SE: Disruption of the Y-box binding protein-1 results in suppression of the epidermal growth factor receptor and HER-2. Cancer Res. 2006, 66: 4872-4879. 10.1158/0008-5472.CAN-05-3561.PubMed
45.
Dhillon J, Astanehe A, Lee C, Fotovati A, Hu K, Dunn SE: The expression of activated Y-box binding protein-1 serine 102 mediates trastuzumab resistance in breast cancer cells by increasing CD44+ cells. Oncogene. 2010, 29: 6294-6300. 10.1038/onc.2010.365.PubMed
46.
Bos JL: ras oncogenes in human cancer: a review. Cancer Res. 1989, 49: 4682-4689.PubMed
47.
Malumbres M, Barbacid M: RAS oncogenes: the first 30 years. Nat Rev Cancer. 2003, 3: 459-465. 10.1038/nrc1097.PubMed
48.
Ramos FJ, Macarulla T, Capdevila J, Elez E, Tabernero J: Understanding the predictive role of K-ras for epidermal growth factor receptor-targeted therapies in colorectal cancer. Clin Colorectal Cancer. 2008, 7 (Suppl 2): S52-S57. 10.3816/CCC.2008.s.008.PubMed
49.
Eng C: K-Ras and sensitivity to EGFR inhibitors in metastatic colorectal cancer. Clin Adv Hematol Oncol. 2008, 6: 174-175.PubMed
50.
Oldham SM, Clark GJ, Gangarosa LM, Coffey RJ, Der CJ: Activation of the Raf-1/MAP kinase cascade is not sufficient for Ras transformation of RIE-1 epithelial cells. Proc Natl Acad Sci USA. 1996, 93: 6924-6928. 10.1073/pnas.93.14.6924.PubMedPubMedCentral
51.
Ciardiello F, Kim N, McGeady ML, Liscia DS, Saeki T, Bianco C, Salomon DS: Expression of transforming growth factor α (TGFα) in breast cancer. Ann Oncol. 1991, 2: 169-182.PubMed
52.
Gangarosa LM, Sizemore N, Graves-Deal R, Oldham SM, Der CJ, Coffey RJ: A raf-independent epidermal growth factor receptor autocrine loop is necessary for Ras transformation of rat intestinal epithelial cells. J Biol Chem. 1997, 272: 18926-18931. 10.1074/jbc.272.30.18926.PubMed
53.
Sizemore N, Cox AD, Barnard JA, Oldham SM, Reynolds ER, Der CJ, Coffey RJ: Pharmacological inhibition of Ras-transformed epithelial cell growth is linked to down-regulation of epidermal growth factor-related peptides. Gastroenterology. 1999, 117: 567-576. 10.1016/S0016-5085(99)70449-X.PubMed
54.
Toulany M, Dittmann K, Krüger M, Baumann M, Rodemann HP: Radioresistance of K-Ras mutated human tumor cells is mediated through EGFR-dependent activation of PI3K-AKT pathway. Radiother Oncol. 2005, 76: 143-150. 10.1016/j.radonc.2005.06.024.PubMed
55.
Grana TM, Sartor CI, Cox AD: Epidermal growth factor receptor autocrine signaling in RIE-1 cells transformed by the Ras oncogene enhances radiation resistance. Cancer Res. 2003, 63: 7807-7814.PubMed
56.
Bernhard EJ, Stanbridge EJ, Gupta S, Gupta AK, Soto D, Bakanauskas VJ, Cerniglia GJ, Muschel RJ, McKenna WG: Direct evidence for the contribution of activated N-ras and K-ras oncogenes to increased intrinsic radiation resistance in human tumor cell lines. Cancer Res. 2000, 60: 6597-6600.PubMed
57.
Kidd AR, Snider JL, Martin TD, Graboski SF, Der CJ, Cox AD: Ras-related small GTPases RalA and RalB regulate cellular survival after ionizing radiation. Int J Radiat Oncol Biol Phys. 2010, 78: 205-212. 10.1016/j.ijrobp.2010.03.023.PubMedPubMedCentral
Metadata
Title
Impact of oncogenic K-RASon YB-1 phosphorylation induced by ionizing radiation
Authors
Mahmoud Toulany
Tim-Andre Schickfluß
Wolfgang Eicheler
Rainer Kehlbach
Birgit Schittek
H Peter Rodemann
Publication date
01-04-2011
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 2/2011
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr2845

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