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Breast Cancer Research
Published in: Breast Cancer Research 4/2011

Open Access 01-08-2011 | Research article

Tribbles homolog 3 denotes a poor prognosis in breast cancer and is involved in hypoxia response

Authors: Marloes Wennemers, Johan Bussink, Blanca Scheijen, 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

Published in: Breast Cancer Research | Issue 4/2011

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Abstract

Introduction

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.

Methods

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.

Results

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.

Conclusions

TRIB3 is independently associated with poor prognosis of breast cancer patients, possibly through its association with tumor cell hypoxia.
Appendix
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Literature
1.
Harris AL: Hypoxia--a key regulatory factor in tumour growth. Nat Rev Cancer. 2002, 2: 38-47. 10.1038/nrc704.CrossRefPubMed
2.
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
3.
Milani M, Harris AL: Targeting tumour hypoxia in breast cancer. Eur J Cancer. 2008, 44: 2766-2773. 10.1016/j.ejca.2008.09.025.CrossRefPubMed
4.
5.
Bos R, Zhong H, Hanrahan CF, Mommers EC, Semenza GL, Pinedo HM, Abeloff MD, Simons JW, van Diest PJ, van der Wall E: Levels of hypoxia-inducible factor-1 alpha during breast carcinogenesis. J Natl Cancer Inst. 2001, 93: 309-314. 10.1093/jnci/93.4.309.CrossRefPubMed
6.
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
7.
Rzymski T, Paantjens A, Bod J, Harris AL: Multiple pathways are involved in the anoxia response of SKIP3 including HuR-regulated RNA stability, NF-kappaB and ATF4. Oncogene. 2008, 27: 4532-4543. 10.1038/onc.2008.100.CrossRefPubMed
8.
Rouschop KM, Wouters BG: Regulation of autophagy through multiple independent hypoxic signaling pathways. Curr Mol Med. 2009, 9: 417-424. 10.2174/156652409788167131.CrossRefPubMed
9.
Ohoka N, Yoshii S, Hattori T, Onozaki K, Hayashi H: TRB3, a novel ER stress-inducible gene, is induced via ATF4-CHOP pathway and is involved in cell death. Embo Journal. 2005, 24: 1243-1255. 10.1038/sj.emboj.7600596.CrossRefPubMedPubMedCentral
10.
Du KY, Herzig S, Kulkarni RN, Montminy M: TRB3: A tribbles homolog that inhibits Akt/PKB activation by insulin in liver. Science. 2003, 300: 1574-1577. 10.1126/science.1079817.CrossRefPubMed
11.
Kiss-Toth E, Bagstaff SM, Sung HY, Jozsa V, Dempsey C, Caunt JC, Oxley KM, Wyllie DH, Polgar T, Harte M, et al: Human tribbles, a protein family controlling mitogen-activated protein kinase cascades. Journal of Biological Chemistry. 2004, 279: 42703-42708. 10.1074/jbc.M407732200.CrossRefPubMed
12.
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
13.
Schwarzer R, Dames S, Tondera D, Klippel A, Kaufmann J: TRB3 is a PI 3-kinase dependent indicator for nutrient starvation. Cellular Signalling. 2006, 18: 899-909. 10.1016/j.cellsig.2005.08.002.CrossRefPubMed
14.
Bowers AJ, Scully S, Boylan JF: SKIP3, a novel Drosophila tribbles ortholog, is overexpressed in human tumors and is regulated by hypoxia. Oncogene. 2003, 22: 2823-2835. 10.1038/sj.onc.1206367.CrossRefPubMed
15.
Ord D, Ord T: Characterization of human NIPK (TRB3, SKIP3) gene activation in stressful conditions. Biochemical and Biophysical Research Communications. 2005, 330: 210-218. 10.1016/j.bbrc.2005.02.149.CrossRefPubMed
16.
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
17.
Miyoshi N, Ishii H, Mimori K, Takatsuno Y, Kim H, Hirose H, Sekimoto M, Doki Y, Mori M: Abnormal expression of TRIB3 in colorectal cancer: a novel marker for prognosis. Br J Cancer. 2009, 101: 1664-1670. 10.1038/sj.bjc.6605361.CrossRefPubMedPubMedCentral
18.
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
19.
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
20.
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
21.
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
22.
Span PN, Bussink J, Manders P, Beex LV, Sweep CG: Carbonic anhydrase-9 expression levels and prognosis in human breast cancer: association with treatment outcome. Br J Cancer. 2003, 89: 271-276. 10.1038/sj.bjc.6601122.CrossRefPubMedPubMedCentral
23.
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
24.
25.
McShane LM, Altman DG, Sauerbrei W, Taube SE, Gion M, Clark GM: Reporting recommendations for tumor marker prognostic studies (REMARK). J Natl Cancer Inst. 2005, 97: 1180-1184. 10.1093/jnci/dji237.CrossRefPubMed
26.
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
27.
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
28.
Raleigh JA, Chou SC, Bono EL, Thrall DE, Varia MA: Semiquantitative immunohistochemical analysis for hypoxia in human tumors. Int J Radiat Oncol Biol Phys. 2001, 49: 569-574. 10.1016/S0360-3016(00)01505-4.CrossRefPubMed
29.
Arteel GE, Thurman RG, Yates JM, Raleigh JA: Evidence that hypoxia markers detect oxygen gradients in liver: pimonidazole and retrograde perfusion of rat liver. Br J Cancer. 1995, 72: 889-895. 10.1038/bjc.1995.429.CrossRefPubMedPubMedCentral
30.
Ljungkvist AS, Bussink J, Rijken PF, Kaanders JH, van der Kogel AJ, Denekamp J: Vascular architecture, hypoxia, and proliferation in first-generation xenografts of human head-and-neck squamous cell carcinomas. Int J Radiat Oncol Biol Phys. 2002, 54: 215-228.CrossRefPubMed
31.
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
32.
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
33.
Rijken PF, Bernsen HJ, van der Kogel AJ: Application of an image analysis system to the quantitation of tumor perfusion and vascularity in human glioma xenografts. Microvasc Res. 1995, 50: 141-153. 10.1006/mvre.1995.1048.CrossRefPubMed
34.
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
35.
Joseph JK, Bunnachak D, Burke TJ, Schrier W: A novel method of inducing and assuring total anoxia during in vitro studies of O2 deprivation injury. J Am Soc Nephrol. 1990, 1: 837-840.PubMed
36.
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970, 227: 680-685. 10.1038/227680a0.CrossRefPubMed
37.
Rademakers SE, Span PN, Kaanders JH, Sweep FC, van der Kogel AJ, Bussink J: Molecular aspects of tumour hypoxia. Mol Oncol. 2008, 2: 41-53. 10.1016/j.molonc.2008.03.006.CrossRefPubMed
38.
Grosshans J, Wieschaus E: A genetic link between morphogenesis and cell division during formation of the ventral furrow in Drosophila. Cell. 2000, 101: 523-531. 10.1016/S0092-8674(00)80862-4.CrossRefPubMed
39.
Mata J, Curado S, Ephrussi A, Rorth P: Tribbles coordinates mitosis and morphogenesis in Drosophila by regulating string/CDC25 proteolysis. Cell. 2000, 101: 511-522. 10.1016/S0092-8674(00)80861-2.CrossRefPubMed
40.
Seher TC, Leptin M: Tribbles, a cell-cycle brake that coordinates proliferation and morphogenesis during Drosophila gastrulation. Current Biology. 2000, 10: 623-629. 10.1016/S0960-9822(00)00502-9.CrossRefPubMed
41.
Sakai S, Ohoka N, Onozaki K, Kitagawa M, Nakanishi M, Hayashi H: Dual Mode of Regulation of Cell Division Cycle 25 A Protein by TRB3. Biol Pharm Bull. 2010, 33: 1112-1116. 10.1248/bpb.33.1112.CrossRefPubMed
42.
Kato S, Du KY: TRB3 modulates C2C12 differentiation by interfering with Akt activation. Biochem Biophys Res Commun. 2007, 353: 933-938. 10.1016/j.bbrc.2006.12.161.CrossRefPubMed
43.
Koo SH, Satoh H, Herzig S, Lee CH, Hedrick S, Kulkarni R, Evans RM, Olefsky J, Montminy M: PGC-1 promotes insulin resistance in liver through PPAR-alpha-dependent induction of TRB-3. Nat Med. 2004, 10: 530-534. 10.1038/nm1044.CrossRefPubMed
44.
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
45.
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
46.
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
47.
Bi M, Naczki C, Koritzinsky M, Fels D, Blais J, Hu N, Harding H, Novoa I, Varia M, Raleigh J, et al: ER stress-regulated translation increases tolerance to extreme hypoxia and promotes tumor growth. EMBO J. 2005, 24: 3470-3481. 10.1038/sj.emboj.7600777.CrossRefPubMedPubMedCentral
48.
Romero-Ramirez L, Cao H, Nelson D, Hammond E, Lee AH, Yoshida H, Mori K, Glimcher LH, Denko NC, Giaccia AJ, et al: XBP1 is essential for survival under hypoxic conditions and is required for tumor growth. Cancer Res. 2004, 64: 5943-5947. 10.1158/0008-5472.CAN-04-1606.CrossRefPubMed
49.
Koumenis C, Wouters BG: "Translating" tumor hypoxia: unfolded protein response (UPR)-dependent and UPR-independent pathways. Mol Cancer Res. 2006, 4: 423-436. 10.1158/1541-7786.MCR-06-0150.CrossRefPubMed
50.
Koritzinsky M, Rouschop KM, van den Beucken T, Magagnin MG, Savelkouls K, Lambin P, Wouters BG: Phosphorylation of eIF2 alpha is required for mRNA translation inhibition and survival during moderate hypoxia. Radiother Oncol. 2007, 83: 353-361. 10.1016/j.radonc.2007.04.031.CrossRefPubMed
Metadata
Title
Tribbles homolog 3 denotes a poor prognosis in breast cancer and is involved in hypoxia response
Authors
Marloes Wennemers
Johan Bussink
Blanca Scheijen
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
Publication date
01-08-2011
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 4/2011
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr2934

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