End-binding protein 1 stimulates paclitaxel sensitivity in breast cancer by promoting its actions toward microtubule assembly and stability

Abstract

Paclitaxel is a microtubule-targeting agent widely used for the treatment of many solid tumors. However, patients show variable sensitivity to this drug, and effective diagnostic tests predicting drug sensitivity remain to be investigated. Herein, we show that the expression of end-binding protein 1 (EB1), a regulator of microtubule dynamics involved in multiple cellular activities, in breast tumor tissues correlates with the pathological response of tumors to paclitaxel-based chemotherapy. In vitro cell proliferation assays reveal that EB1 stimulates paclitaxel sensitivity in breast cancer cell lines. Our data further demonstrate that EB1 increases the activity of paclitaxel to cause mitotic arrest and apoptosis in cancer cells. In addition, microtubule binding affinity analysis and polymerization/depolymerization assays show that EB1 enhances paclitaxel binding to microtubules and stimulates the ability of paclitaxel to promote microtubule assembly and stabilization. These findings thus reveal EB1 as a critical regulator of paclitaxel sensitivity and have important implications in breast cancer chemotherapy.

Keywords

breast cancer / paclitaxel / microtubule / cell proliferation / apoptosis / end-binding protein 1 / chemotherapy

References

[1] Akhmanova A, Steinmetz MO (2008) Tracking the ends: a dynamic protein network controls the fate of microtubule tips. Nat Rev Mol Cell Biol9: 309322

[2] Amos LA, Schlieper D (2005) Microtubules and maps. Adv Protein Chem71: 257298

[3] Asakawa K, Toda T (2006) Cooperation of EB1-Mal3 and the Bub1 spindle checkpoint. Cell Cycle5: 2730

[4] Bieling P, Laan L, Schek H, Munteanu EL, Sandblad L, Dogterom M, Brunner D, Surrey T (2007) Reconstitution of a microtubule plusend tracking system in vitro. Nature450: 11001105

[5] Blagosklonny MV, Demidenko ZN, Giovino M, Szynal C, Donskoy E, Herrmann RA, Barry JJ, Whalen AM (2006) Cytostatic activity of paclitaxel in coronary artery smooth muscle cells is mediated through transient mitotic arrest followed by permanent postmitotic arrest: comparison with cancer cells. Cell Cycle5: 15741579

[6] Bu W, Su LK (2001) Regulation of microtubule assembly by human EB1 family proteins. Oncogene20: 31853192

[7] Dong X, Liu F, Sun L, Liu M, Li D, Su D, Zhu Z, Dong JT, Fu L, Zhou J (2010) Oncogenic function of microtubule end-binding protein 1 in breast cancer. J Pathol220: 361369

[8] Dragestein KA, van Cappellen WA, van Haren J, Tsibidis GD, Akhmanova A, Knoch TA, Grosveld F, Galjart N (2008) Dynamic behavior of GFP-CLIP-170 reveals fast protein turnover on microtubule plus ends. J Cell Biol180: 729737

[9] Dumontet C, Jordan MA (2010) Microtubule-binding agents: a dynamic field of cancer therapeutics. Nat Rev Drug Discov9: 790803

[10] Galjart N (2010) Plus-end-tracking proteins and their interactions at microtubule ends. Curr Biol20: R528R537

[11] Honnappa S, Gouveia SM, Weisbrich A, Damberger FF, Bhavesh NS, Jawhari H, Grigoriev I, van Rijssel FJ, Buey RM, Lawera A (2009) An EB1-binding motif acts as a microtubule tip localization signal. Cell138: 366376

[12] Huo L, Li D, Sun L, Liu M, Shi X, Sun X, Li J, Dong B, Dong X, Zhou J (2011) Tat acetylation regulates its actions on microtubule dynamics and apoptosis in T lymphocytes. J Pathol223: 2836

[13] Ikui AE, Yang CP, Matsumoto T, Horwitz SB (2005) Low concentrations of taxol cause mitotic delay followed by premature dissociation of p55CDC from Mad2 and BubR1 and abrogation of the spindle checkpoint, leading to aneuploidy. Cell Cycle4: 13851388

[14] Jiang K, Akhmanova A (2011) Microtubule tip-interacting proteins: a view from both ends. Curr Opin Cell Biol23: 94101

[15] Jordan MA, Wilson L (2004) Microtubules as a target for anticancer drugs. Nat Rev Cancer4: 253265

[16] Komlodi-Pasztor E, Sackett D, Wilkerson J, Fojo T (2011) Mitosis is not a key target of microtubule agents in patient tumors. Nat Rev Clin Oncol8: 244250

[17] Kronja I, Kruljac-Letunic A, Caudron-Herger M, Bieling P, Karsenti E (2009) XMAP215-EB1 interaction is required for proper spindle assembly and chromosome segregation in Xenopus egg extract. Mol Biol Cell20: 26842696

[18] Li D, Xie S, Ren Y, Huo L, Gao J, Cui D, Liu M, Zhou J (2011) Microtubule-associated deacetylase HDAC6 promotes angiogenesis by regulating cell migration in an EB1-dependent manner. Protein Cell2: 150160

[19] Liu M, Yu H, Huo L, Liu J, Li M, Zhou J (2008) Validating the mitotic kinesin Eg5 as a therapeutic target in pancreatic cancer cells and tumor xenografts using a specific inhibitor. Biochem Pharmacol76: 169178

[20] Mohan R, Katrukha EA, Doodhi H, Smal I, Meijering E, Kapitein LC, Steinmetz MO, Akhmanova A (2013) End-binding proteins sensitize microtubules to the action of microtubule-targeting agents. Proc Natl Acad Sci USA110: 89008905

[21] Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T, Clark L, Bayani N, Coppe JP, Tong F (2006) A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell10: 515527

[22] Quinn JE, James CR, Stewart GE, Mulligan JM, White P, Chang GK, Mullan PB, Johnston PG, Wilson RH, Harkin DP (2007) BRCA1 mRNA expression levels predict for overall survival in ovarian cancer after chemotherapy. Clin Cancer Res13: 74137420

[23] Rouzier R, Rajan R, Wagner P, Hess KR, Gold DL, Stec J, Ayers M, Ross JS, Zhang P, Buchholz TA (2005) Microtubuleassociated protein tau: a marker of paclitaxel sensitivity in breast cancer. Proc Natl Acad Sci USA102: 83158320

[24] Schuyler SC, Pellman D (2001) Microtubule plus-end-tracking proteins: The end is just the beginning. Cell105: 421424

[25] Strickland LI, Wen Y, Gundersen GG, Burgess DR (2005) Interaction between EB1 and p150glued is required for anaphase astral microtubule elongation and stimulation of cytokinesis. Curr Biol15: 22492255

[26] Sun L, Gao J, Dong X, Liu M, Li D, Shi X, Dong JT, Lu X, Liu C, Zhou J (2008) EB1 promotes Aurora-B kinase activity through blocking its inactivation by protein phosphatase 2A. Proc Natl Acad Sci USA105: 71537158

[27] Sun X, Shi X, Liu M, Li D, Zhang L, Liu X, Zhou J (2011) Mdp3 is a novel microtubule-binding protein that regulates microtubule assembly and stability. Cell Cycle10: 39293937

[28] Sun X, Li D, Yang Y, Ren Y, Li J, Wang Z, Dong B, Liu M, Zhou J (2012) Microtubule-binding protein CLIP-170 is a mediator of paclitaxel sensitivity. J Pathol226: 666673

[29] Sun X, Li F, Dong B, Suo S, Liu M, Li D, Zhou J (2013) Regulation of tumor angiogenesis by the microtubule-binding protein CLIP-170. Protein Cell4: 266276

[30] Tommasi S, Mangia A, Lacalamita R, Bellizzi A, Fedele V, Chiriatti A, Thomssen C, Kendzierski N, Latorre A, Lorusso V (2007) Cytoskeleton and paclitaxel sensitivity in breast cancer: the role of beta-tubulins. Int J Cancer120: 20782085

[31] Veitia R, David S, Barbier P, Vantard M, Gounon P, Bissery MC, Fellous A (2000) Proteolysis of microtubule associated protein 2 and sensitivity of pancreatic tumours to docetaxel. Brit J Cancer83: 544549

[32] Vichai V, Kirtikara K (2006) Sulforhodamine B colorimetric assay for cytotoxicity screening. Nat Protoc1: 11121116

[33] Vitre B, Coquelle FM, Heichette C, Garnier C, Chretien D, Arnal I (2008) EB1 regulates microtubule dynamics and tubulin sheet closure in vitro. Nat Cell Biol10: 415421

[34] Wagner P, Wang B, Clark E, Lee H, Rouzier R, Pusztai L (2005) Microtubule associated protein (MAP)-tau: a novel mediator of paclitaxel sensitivity in vitro and in vivo. Cell Cycle4: 11491152

[35] Wang H, Liu B, Zhang C, Peng G, Liu M, Li D, Gu F, Chen Q, Dong JT, Fu L (2009) Parkin regulates paclitaxel sensitivity in breast cancer via a microtubule-dependent mechanism. J Pathol218: 7685

[36] Wen Y, Eng CH, Schmoranzer J, Cabrera-Poch N, Morris EJS, Chen M, Wallar BJ, Alberts AS, Gundersen GG (2004) EB1 and APC bind to mDia to stabilize microtubules downstream of Rho and promote cell migration. Nat Cell Biol6: 820830

[37] Zovko S, Abrahams JP, Koster AJ, Galjart N, Mommaas AM (2008) Microtubule plus-end conformations and dynamics in the periphery of interphase mouse fibroblasts. Mol Biol Cell19: 31383146

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