The online version of this article (doi:10.1186/bcr2934) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
MW performed laboratory experiments, data analysis and participated in the experimental design and drafting of the manuscript. JB participated in the design and coordination of the research and assisted in xenograft assays. BS contributed to the interpretation of the data and critical revised the manuscript. IDN participated in the immunohistochemical stainings and the interpretation of these data. HWML participated in the collection of the patient material and data. JAR and MAV participated in the collection of patient material containing pimonidazole. JJTMH participated in RT-qPCR experiments and interpretation of the results. KMR carried out the MCF-7 knockdown experiments and critically revised the manuscript. FCGJS participated in the design and coordination of the research and assisted in the patient material collection. PNS participated in the design and coordination of the research, performed the statistical analysis and helped to draft the manuscript. All authors read and approved the final manuscript.
Hypoxia in solid tumors is associated with treatment resistance, resulting in poor prognosis. Tribbles homolog 3 (TRIB3) is induced during hypoxia and is involved in multiple cellular pathways involved in cell survival. Here, we investigated the role of TRIB3 in breast cancer.
TRIB3 mRNA expression was measured in breast tumor tissue from 247 patients and correlated with clinicopathological parameters and clinical outcome. Furthermore, we studied TRIB3 expression regulation in cell lines, xenografts tissues and human breast cancer material using Reverse transcriptase, quantitative polymerase chain reaction (RT-qPCR) and immunohistochemical staining. Finally, the effect of small interfering RNA (siRNA) mediated TRIB3 knockdown on hypoxia tolerance was assessed.
Breast cancer patients with low, intermediate or high TRIB3 expression exhibited a mean disease free survival (DFS) of 80 (95% confidence interval [CI] = 74 to 86), 74 (CI = 67 to 81), and 63 (CI = 55 to 71) months respectively (P = .002, Mantel-Cox log-rank). The prognostic value of TRIB3 was limited to those patients that had received radiotherapy as part of their primary treatment (n = 179, P = .005) and remained statistically significant after correction for other clinicopathological parameters (DFS, Hazard Ratio = 1.90, CI = 1.17 to 3.08, P = .009). In breast cell lines TRIB3 expression was induced by hypoxia, nutrient starvation, and endoplasmic reticulum stress in an hypoxia inducible factor 1 (HIF-1) independent manner. TRIB3 induction after hypoxia did not increase with decreasing oxygen levels. In breast tumor xenografts and human breast cancer tissues TRIB3 co-localized with the hypoxic cell marker pimonidazole. The induction of TRIB3 by hypoxia was shown to be regulated via the PERK/ATF4/CHOP pathway of the unfolded protein response and knockdown of TRIB3 resulted in a dose-dependent increase in hypoxia sensitivity.
TRIB3 is independently associated with poor prognosis of breast cancer patients, possibly through its association with tumor cell hypoxia.
Authors’ original file for figure 113058_2011_2734_MOESM1_ESM.pdf
Authors’ original file for figure 213058_2011_2734_MOESM2_ESM.pdf
Authors’ original file for figure 313058_2011_2734_MOESM3_ESM.pdf
Authors’ original file for figure 413058_2011_2734_MOESM4_ESM.pdf
Authors’ original file for figure 513058_2011_2734_MOESM5_ESM.pdf
Hockel M, Schlenger K, Aral B, Mitze M, Schaffer U, Vaupel P: Association between tumor hypoxia and malignant progression in advanced cancer of the uterine cervix. Cancer Res. 1996, 56: 4509-4515. PubMed
Generali D, Berruti A, Brizzi MP, Campo L, Bonardi S, Wigfield S, Bersiga A, Allevi G, Milani M, Aguggini S, et al: Hypoxia-inducible factor-1 alpha expression predicts a poor response to primary chemoendocrine therapy and disease-free survival in primary human breast cancer. Clin Cancer Res. 2006, 12: 4562-4568. 10.1158/1078-0432.CCR-05-2690. CrossRefPubMed
Ord D, Meerits K, Ord T: TRB3 protects cells against the growth inhibitory and cytotoxic effect of ATF4. Experimental Cell Research. 2007, 3 (13): 3556-3567. CrossRef
Salazar M, Carracedo A, Salanueva IJ, Hernandez-Tiedra S, Lorente M, Egia A, Vazquez P, Blazquez C, Torres S, Garcia S, et al: Cannabinoid action induces autophagy-mediated cell death through stimulation of ER stress in human glioma cells. Journal of Clinical Investigation. 2009, 119: 1359-1372. 10.1172/JCI37948. CrossRefPubMedPubMedCentral
Xu JM, Lv S, Qin Y, Shu F, Xu YJ, Chen J, Xu BE, Sun XQ, Wu J: TRB3 interacts with CtIP and is overexpressed in certain cancers. Biochimica Et Biophysica Acta-General Subjects. 2007, 1770: 273-278. 10.1016/j.bbagen.2006.09.025. CrossRef
Chi JT, Wang Z, Nuyten DS, Rodriguez EH, Schaner ME, Salim A, Wang Y, Kristensen GB, Helland A, Borresen-Dale AL, et al: Gene expression programs in response to hypoxia: cell type specificity and prognostic significance in human cancers. PLoS Med. 2006, 3: e47-10.1371/journal.pmed.0030047. CrossRefPubMedPubMedCentral
Chia SK, Wykoff CC, Watson PH, Han C, Leek RD, Pastorek J, Gatter KC, Ratcliffe P, Harris AL: Prognostic significance of a novel hypoxia-regulated marker, carbonic anhydrase IX, in invasive breast carcinoma. J Clin Oncol. 2001, 19: 3660-3668. PubMed
Davies MP, Barraclough DL, Stewart C, Joyce KA, Eccles RM, Barraclough R, Rudland PS, Sibson DR: Expression and splicing of the unfolded protein response gene XBP-1 are significantly associated with clinical outcome of endocrine-treated breast cancer. Int J Cancer. 2008, 123: 85-88. 10.1002/ijc.23479. CrossRefPubMed
Nicholson RI, Hutcheson IR, Britton D, Knowlden JM, Jones HE, Harper ME, Hiscox SE, Barrow D, Gee JM: Growth factor signalling networks in breast cancer and resistance to endocrine agents: new therapeutic strategies. J Steroid Biochem Mol Biol. 2005, 93: 257-262. 10.1016/j.jsbmb.2004.12.006. CrossRefPubMed
Span PN, Waanders E, Manders P, Heuvel JJ, Foekens JA, Watson MA, Beex LV, Sweep FC: Mammaglobin is associated with low-grade, steroid receptor-positive breast tumors from postmenopausal patients, and has independent prognostic value for relapse-free survival time. J Clin Oncol. 2004, 22: 691-698. 10.1200/JCO.2004.01.072. CrossRefPubMed
Sweep CG, Geurts-Moespot J: EORTC external quality assurance program for ER and PgR measurements: trial 1998/1999. European Organisation for Research and Treatment of Cancer. Int J Biol Markers. 2000, 15: 62-69. PubMed
Wijffels KI, Kaanders JH, Marres HA, Bussink J, Peters HP, Rijken MSPF, van den Hoogen FJ, de Wilde PC, van der Kogel AJ: Patterns of proliferation related to vasculature in human head-and-neck carcinomas before and after transplantation in nude mice. Int J Radiat Oncol Biol Phys. 2001, 51: 1346-1353. 10.1016/S0360-3016(01)02605-0. CrossRefPubMed
Rouschop KM, van den Beucken T, Dubois L, Niessen H, Bussink J, Savelkouls K, Keulers T, Mujcic H, Landuyt W, Voncken JW, et al: The unfolded protein response protects human tumor cells during hypoxia through regulation of the autophagy genes MAP1LC3B and ATG5. J Clin Invest. 2010, 120: 127-141. 10.1172/JCI40027. CrossRefPubMed
Ho KC, Leach JK, Eley K, Mikkelsen RB, Lin PS: A simple method of producing low oxygen conditions with oxyrase for cultured cells exposed to radiation and tirapazamine. Am J Clin Oncol. 2003, 26: e86-91. PubMed
Joseph JK, Bunnachak D, Burke TJ, Schrier W: A novel method of inducing and assuring total anoxia during in vitro studies of O 2 deprivation injury. J Am Soc Nephrol. 1990, 1: 837-840. PubMed
Prudente S, Hribal ML, Flex E, Turchi F, Morini E, De Cosmo S, Bacci S, Tassi V, Cardellini M, Lauro R, et al: The functional Q84R polymorphism of mammalian tribbles homolog TRB3 is associated with insulin resistance and related cardiovascular risk in Caucasians from Italy. Diabetes. 2005, 54: 2807-2811. 10.2337/diabetes.54.9.2807. CrossRefPubMed
Corcoran CA, Luo X, He Q, Jiang C, Huang Y, Sheikh MS: Genotoxic and endoplasmic reticulum stresses differentially regulate TRB3 expression. Cancer biology & therapy. 2005, 4: 1063-1067. 10.4161/cbt.4.10.2205. CrossRef
Wasef SZY, Robinson KA, Berkaw MN, Buse MG: Glucose, dexamethasone, and the unfolded protein response regulate TRB3 mRNA expression in 3T3-L1 adipocytes and L6 myotubes. American Journal of Physiology-Endocrinology and Metabolism. 2006, 291: E1274-E1280. 10.1152/ajpendo.00117.2006. CrossRef
- Tribbles homolog 3 denotes a poor prognosis in breast cancer and is involved in hypoxia response
Iris D Nagtegaal
Hanneke WM van Laarhoven
James A Raleigh
Mahesh A Varia
Joop JTM Heuvel
Kasper M Rouschop
Fred CGJ Sweep
Paul N Span
- BioMed Central
Neu im Fachgebiet Onkologie
Mail Icon II