Abstract
Critics have suggested that neoadjuvant chemotherapy (NACT) followed by interval debulking may select for resistant clones or cancer stem cells when compared to primary cytoreduction. β-tubulins are chemotherapeutic targets of taxanes and epothilones. Class III β-tubulin overexpression has been linked to chemoresistance and hypoxia. Herein, we describe changes in class III β-tubulin in patients with advanced ovarian carcinoma in response to NACT, in relationship to clinical outcome, and between patients who underwent NACT versus primary debulking; we characterize in vitro chemosensitivity to paclitaxel/patupilone of cell lines established from this patient population, and class III β-tubulin expression following repeated exposure to paclitaxel. Using immunohistochemistry, we observed among 22 paired specimens obtained before/after NACT decreased expression of class III β-tubulin following therapy within stroma (p = 0.07), but not tumor (p = 0.63). Poor median overall survival was predicted by high levels of class III β-tubulin in both tumor (HR 3.66 [1.11,12.05], p = 0.03) and stroma (HR 4.53 [1.28,16.1], p = 0.02). Class III β-tubulin expression by quantitative-real-time-polymerase-chain-reaction was higher among patients who received NACT (n = 12) compared to primary cytoreduction (n = 14) (mean ± SD fold-change: 491.2 ± 115.9 vs. 224.1 ± 55.66, p = 0.037). In vitro subculture with paclitaxel resulted in class III β-tubulin upregulation, however, cell lines that overexpressed class III β-tubulin remained sensitive to patupilone. Overexpression of class III β-tubulin in patients dispositioned to NACT may thus identify an intrinsically aggressive phenotype, and predict poor overall survival and paclitaxel resistance. Decreases in stromal expression may represent normalization of the tumor microenvironment following therapy. Epothilones warrant study for patients who have received neoadjuvant carboplatin and paclitaxel.
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Abbreviations
- NACT:
-
Neoadjuvant chemotherapy
- FIGO:
-
Fédération Internationale de Gynécologie et d’Obstétrique
- qRT-PCR:
-
Quantitative real time polymerase chain reaction
- OSPC:
-
Ovarian serous papillary carcinoma
- USPC:
-
Uterine serous papillary carcinoma
- OS:
-
Overall survival
- ΔIHC:
-
Change in immunohistochemistry score
References
Vergote I, Tropé CG, Amant F, Kristensen GB, Ehlen T, Johnson N, Verheijen RHM, van der Burg MEL, Lacave AJ, Benedetti-Panici P, Kenter GG, Casado A, Mendiola C, Coens C, Verleye L, Stuart GC, Pecorelli S, Reed NS (2010) Neoadjuvant chemotherapy or primary surgery in stage IIIC or IV ovarian cancer. New Engl J Med 363:943–953
Kang S, Nam BH (2009) Does neoadjuvant chemotherapy increase optimal cytoreduction rate in advanced ovarian cancer? Meta-analysis of 21 studies. Ann Surg Oncol 16:2315–2320
Crawford SC, Vasey PA, Paul J, Hay A, Kaye SB (2005) Does aggressive surgery only benefit patients with less advanced cancer? Results from an international comparison within the SCOTROC-1 Trial. J Clin Oncol 23:8802–8811
Glasgow MA, Yu H, Rutherford TJ, Azodi M, Silasi DA, Santin AD, Schwartz PE (2013) Neoadjuvant chemotherapy (NACT) is an effective way of managing elderly women with advanced stage ovarian cancer (FIGO Stage IIIC and IV). J Surg Oncol 107(2):195–200
Menczer J, Usviatzov I, Ben-Shem E, Golan A, Levy T (2011) Neoadjuvant chemotherapy in ovarian, primary peritoneal and tubal carcinoma: can imaging results prior to interval debulking predict survival? J Gynecol Oncol 22(3):183–187
Bristow RE, Chi DS (2006) Platinum-based neoadjuvant chemotherapy and interval surgical cytoreduction for advanced ovarian cancer: a meta-analysis. Gynecol Oncol 103:1070–1076
Raspaglio G, Fillipetti F, Prislei F, Penci R, De Maria I, Cicchillitti K, Mozzetti S, Scambia G, Ferlini C (2008) Hypoxia induces class III beta-tubulin gene expression by HIF-1 alpha binding to its 3′ flanking region. Gene 409(1–2):100–108
Magnani M, Ortuso F, Soro S, Alcaro S, Tramontano A, Botta M (2006) The β-I/β-III- tubulin isoforms and their complexes with antimitotic agent: docking and molecular dynamics studies. FEBS J 273:3301–3310
De Donato M, Mariani M, Petrella L, Martinelli E, Zannoni GF, Vellone V, Ferrandina G, Shahabi S, Scambia G, Ferlini C (2012) Class III β-tubulin and the cytoskeletal gateway for drug resistance in ovarian cancer. J Cell Physiol 227:1034–1041
Ferlini C, Raspaglio G, Cicchillitti L, Mozzetti S, Prislei S, Bartollino S, Scambia G (2007) Looking at drug resistance mechanisms for microtubule interacting drugs: does TUBB3 work? Curr Cancer Drug Targets 7(8):704–712
Sève P, Dumontet C, Reiman T (2010) Role of tubulin-β-III in predicting chemoresistance in non-small cell lung cancer. Lung Cancer 67(2):136–143
Tommasi S, Mangia A, Lacalamita R, Bellizzi A, Fedele V, Chiriatti A, Thomssen C, Kendzierski N, Latorre A, Lorusso V, Schittulli F, Zito F, Kavallaris M, Paradiso A (2007) Cytoskeleton and paclitaxel sensitivity in breast cancer: the role of β-tubulins. Int J Cancer 120:2078–2085
Mariani M, Zannoni G, Sioletic S, Sieber S, Martino C, Martinelli E, Coco C, Scambia G, Shahabi S, Ferlini C (2012) Gender influences the Class III and V beta-tubulin ability to predict poor outcome in colorectal cancer. Clin Cancer Res 18(10):2964–2975
Sève P, Reiman T, Lai R, Hanson J, Santos C, Johnson L, Dabbagh L, Sawyer M, Dumontet C, Mackey JR (2007) Class III β-tubulin is a marker of paclitaxel resistance in carcinomas of unknown primary site. Cancer Chemother Pharmacol 60:27–34
Bollag DM, McQueney PA, Zhu J, Hensens O, Koupal L, Liesch J, Goetz M, Lazarides E, Woods CM (1995) Epothilones, a new class of microtubule-stabilizing agents with a taxol-like mechanism of action. Cancer Res 55:2325–2333
Tavassoli FA, Devilee P (2003) World Health Organization: Tumours of the Breast and Female Genital Organs. IARC Press, Lyon
El-Sahwi K, Bellone S, Cocco E, Cargnelutti M, Casagrande F, Bellone M, Abu-Khalaf M, Buza N, Tavassoli FA, Hui P, Silasi DA, Azodi M, Schwartz PE, Rutherford TJ, Pecorelli S, Santin AD (2010) In vitro activity of pertuzumab in combination with trastuzumab in uterine serous papillary adenocarcinoma. Br J Cancer 102(1):134–143
Tassi RA, Bignotti E, Rossi E (2007) Overexpression of mammaglobin B in epithelial ovarian carcinomas. Gynecol Oncol 105:578–585
Carrara L, Guzzo F, Roque DM, Bellone S, Cocco E, Sartori E, Pecorelli S, Schwartz PE, Rutherford TJ, Santin AD (2012) Differential in vitro sensitivity to patupilone versus paclitaxel in uterine and ovarian carcinosarcoma cell lines is linked to tubulin-beta-III expression. Gynecol Oncol 125(1):231–236
Mozzetti S, Ferlini C, Concolino P, Filippetti F, Raspaglio G, Prislei S, Gallo D, Martinelli E, Ranelletti F, Ferrandina G, Scambia G (2005) Class III β-tubulin overexpression is a prominent mechanism of paclitaxel resistance in ovarian cancer patients. Clin Cancer Res 11:298–305
Roque DM, Bellone S, English DP, Buza N, Cocco E, Gasparrini S, Bortolomai I, Ratner E, Silasi DA, Azodi M, Rutherford TJ, Schwartz PE, Santin AD (2013) Tubulin-β-III overexpression by uterine serous carcinomas is a marker for poor overall survival after platinum/taxane chemotherapy and sensitivity to epothilones. Cancer. doi:10.1002/cncr.28017
Roque DM, Bellone S, Buza N, Romani C, Cocco E, Bignotti E, Ravaggi A, Rutherford TJ, Schwartz PE, Pecorelli S, Santin AD (2013) Class III β-tubulin overexpression in ovarian clear cell and serous carcinoma as a marker for poor overall survival after platinum/taxane chemotherapy and sensitivity to patupilone. Am J Obstet Gynecol. doi:10.1016/ajog.2013.04.017
Umezu T, Shibata K, Kajiyama H, Terauchi M, Ino K, Nawa A, Kikkawa F (2008) Taxol resistance among the different histological subtypes of ovarian cancer may be associated with the expression of class III beta-tubulin. Int J Gynecol Pathol 27(2):207–212
Tian Q, Stepaniants SB, Mao M, Weng L, Feetham MC, Doyle MJ, Yi EC, Dai H, Thorsson V, Eng J, Goodlett D, Berger JP, Gunter B, Linseley PS, Stoughton RB, Aebersold R, Collins SJ, Hanlon WA, Hood LE (2004) Integrated genomic and proteomic analyses of gene expression in mammalian cells. Mol Cell Proteomics 3:960–969
Vergote I, De Wever I, Tjalma W, Van Gramberen M, Decloedt J, van Dam P (1998) Neoadjuvant chemotherapy or primary debulking surgery in advanced ovarian carcinoma: a retrospective analysis of 285 patients. Gynecol Oncol 71:431–436
Zivanovic O, Eisenhauer EL, Zhou Q, Iasonos A, Sabbatini P, Sonoda Y, Abu-Rustum NR, Barakat RR, Chi DS (2008) The impact of bulky upper abdominal disease cephalad to the greater omentum on surgical outcome for stage IIIC epithelial ovarian, fallopian tube, and primary peritoneal cancer. Gynecol Oncol 108:287–292
Aletti GD, Dowdy SC, Podratz KC, Cliby WA (2006) Surgical treatment of diaphragm disease correlates with improved survival in optimally debulked advanced stage ovarian cancer. Gynecol Oncol 100:283–287
Bristow RE, Tomacruz RS, Armstrong DK, Trimble EL, Montz FJ (2002) Survival effect of maximal cytoreductive surgery for advanced ovarian carcinoma during the platinum era: a meta-analysis. J Clin Oncol 20:1248–1259
Chi DS, Eisenhauer EL, Lang J, Huh J, Haddad L, Abu-Rustum NR, Sonoda Y, Levine DA, Hensley M, Barakat RR (2006) What is the optimal goal of primary cytoreductive surgery for bulky stage IIIC epithelial ovarian carcinoma (EOC)? Gynecol Oncol 103:559–564
du Bois A, Reuss A, Pujade-Lauraine E, Harter P, Ray-Coquard I, Pfisterer J (2009) Role of surgical outcome as prognostic factor in advanced epithelial ovarian cancer: a combined exploratory analysis of 3 prospectively randomized phase 3 multi- center trials: by the Arbeitsgemeinschaft Gynaekologische Onkologie Studiengruppe Ovarialkarzinom (AGO-OVAR) and the Groupe d’Investigateurs Nationaux Pour les Etudes des Cancers de l’Ovaire (GINECO). Cancer 115:1234–1244
Kavallaris M, Kuo DY, Burkhart CA, Regl DL, Norris MD, Haber M, Horwitz SB (1997) Taxol-resistant epithelial ovarian tumors are associated with altered expression of specific beta-tubulin isotypes. J Clin Invest 100(5):1282–1293
Mozzetti S, Ferlini C, Concolino P, Filippetti, Raspaglio G, Prislei S, Gallo D, Martinelli E, Ranelletti FO, Ferrandina G, Scambia G (2005) Class III beta-tubulin overexpression is a prominent mechanism of paclitaxel resistance in ovarian cancer patients. Clin Cancer Res 11(1):298–305
Mariani M, Shahabi S, Sieber S, Scambia G, Ferlini C (2011) Class III β-tubulin (TUBB3): more than a biomarker in solid tumors? Curr Mol Med 11:726–731
Thongwatchara P, Promwikorn W, Srisomsap C, Chokchaichamnankit D, Boonyaphiphat P, Thongsuksai P (2011) Differential protein expression in primary breast cancer and match axillary node metastasis. Oncol Rep 26(1):185–189
Kang CH, Jang BG, Kim D-W, Chung DH, Kim YT, Jheon S, Sung S-W, Hyun J (2009) Differences in the expression profiles of excision repair cross complementation group 1, X-ray repair crosscomplementation group 1, and β-III tubulin between primary non-small cell lung cancer and metastatic lymph nodes and the significance in mid-term survival. J Thoracic Oncol 4(11):1307–1312
van Diest PJ, van Dam P, Henzen-Logmans SC, Berns E, van der Burg ME, Green J, Vergote I (1997) A scoring system for immunohistochemical staining: concensus report of the task force for basic research of the EORTC-GCCG. J Clin Pathol 50:801–804
Ferrandina G, Zannoni GF, Martinelli E, Paglia A, Gallotta V, Mozzetti S, Scambia G, Ferlini C (2006) Class III β-Tubulin overexpression is a marker of poor clinical outcome in advanced ovarian cancer patients. Clin Cancer Res 12:2774–2789
Chefetz I, AlveroAB Holberg JC, Lebowitz N, Craveiro V, Yang-Hatwich Y, Yin G, Squillace L, Sorteras M, Aldo P, Mor G (2013) TLR2 enhances ovarian cancer stem cell self-renewal and promotes tumor repair and recurrence. Cell Cycle 12(3):511–521
Ferlini C, Raspaglio G, Mozzetti S, Cicchillitti L, Filippetti F, Gallo D, Fattorusso C, Campiani G, Scambia G (2005) The seco-taxane IDN5390 is able to target class III β-tubulin and to overcome paclitaxel resistance. Cancer Res 65:2397–2405
Ranganathan S, Benetatos CA, Colarusso PJ, Dexter DW, Hudes GR (1998) Altered β-tubulin isotype expression in paclitaxel-resistant human prostate carcinoma cells. Br J Cancer 77:562–566
Del Carmen MG, Birrer M, Shorge JO (2012) Uterine papillary serous cancer: a review of the literature. Gynecol Oncol 127:652–661
Moller KA, Gehrig PA, Van Le L, Secord AA, Schorge J (2004) The role of optimal debulking in advanced stage serous carcinoma of the uterus. Gynecol Oncol 94(1):170–174
Resnik E, Taxy JB (1996) Neoadjuvant chemotherapy in uterine papillary serous carcinoma. Gynecol Oncol 62(1):123–127
Le TD, Yamada SD, Rutgers JL, DiSaia PJ (1999) Complete response of a stage IV uterine papillary serous carcinoma to neoadjuvant chemotherapy with Taxol and carboplatin. Gynecol Oncol 73(3):461–463
Gan PP, Pasquier E, Kavallaris M (2007) Class III tubulin mediates sensitivity to chemotherapeutic drugs in non-small cell lung cancer. Cancer Res 67:9356–9936
Gan PP, McCarroll JA, Byrne FL, Garner J, Kavallaris M (2011) Specific-b-III tubulin isotypes can functionally enhance or diminish epothilone B sensitivity in non-small cell lung cancer cells. PLoS One 6(6):e21717
Pusztai L (2007) Markers predicting clinical benefit in breast cancer from microtubule-targeting agents. Ann Oncol Suppl 12:xii15–20
De Geest K, Blessing JA, Morris RT, Monk BJ, Zweizig SL, Matei D, Muller CY, Richards WE (2009) Phase II clinical trial of ixabepilone in patients with recurrent or persistent platinum- and taxane-resistant ovarian or primary peritoneal cancer: a gynecologic oncology group study. J Clin Oncol 28:149–153
Dizon DS, Blessing JA, McMeekin DS, Sharma SK, DiSilvestro P, Alvarez RD (2009) Phase II trial of ixabepilone as second-line treatment in advanced endometrial cancer: gynecologic Oncology Group Trial 129-P. J Clin Oncol 27:3104–3108
NIH Clinical Trials. Evaluation of carboplatin/paclitaxel/bevacizumab in the treatment of advanced stage endometrial carcinoma. http://clinicaltrials.gov/ct2/show/ NCT00513786. Accessed 02 January 2013
Vansteenkiste J, Lara PN Jr, Le Chevalier T, Breton JL, Bonomi P, Sandler AB, Socinski MA, Delbaldo C, McHenry B, Lebwohl D, Peck R, Edelman MJ (2007) Phase II clinical trial of the epothilone B analog, ixabepilone, in patients with non-small cell lung caner whose tumors have failed first-line platinum-based chemotherapy. J Clin Oncol 25(23):3448–3455
Spigel DR, Anthony Greco F, Waterhouse DM, Shipley DL, Zubkus JD, Bury MJ, Webb CD, Hart LL, Gian VG, Infante JR, Burris HA 3rd, Hainsworth JD (2012) Phase II trial of ixabepilone and carboplatin with or without bevacizumab in patients with previously untreated advanced non-small cell lung cancer. Lung Cancer 78(1):70–75
McMeekin S, Patel R, Verschraegen C, Celano P, Burke J 2nd, Plaxe S, Ghatage P, Giurescu M, Stredder C, ang Y, Schmelter T (2012) Phase I/II study of sagopilone (ZK-EPO) plus carboplatin in women with recurrent platinum-sensitive ovarian cancer. Br J Cancer 106(1):70–76
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Roque, D.M., Buza, N., Glasgow, M. et al. Class III β-tubulin overexpression within the tumor microenvironment is a prognostic biomarker for poor overall survival in ovarian cancer patients treated with neoadjuvant carboplatin/paclitaxel. Clin Exp Metastasis 31, 101–110 (2014). https://doi.org/10.1007/s10585-013-9614-5
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DOI: https://doi.org/10.1007/s10585-013-9614-5