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Breast Cancer Research
Published in: Breast Cancer Research 1/2013

Open Access 01-02-2012 | Research article

Small interfering RNA library screen identified polo-like kinase-1 (PLK1) as a potential therapeutic target for breast cancer that uniquely eliminates tumor-initiating cells

Authors: Kaiji Hu, Jennifer H Law, Abbas Fotovati, Sandra E Dunn

Published in: Breast Cancer Research | Issue 1/2013

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Abstract

Introduction

Triple-negative breast cancer (TNBC) high rate of relapse is thought to be due to the presence of tumor-initiating cells (TICs), molecularly defined as being CD44high/CD24-/low. TICs are resilient to chemotherapy and radiation. However, no currently accepted molecular target exists against TNBC and, moreover, TICs. Therefore, we sought the identification of kinase targets that inhibit TNBC growth and eliminate TICs.

Methods

A genome-wide human kinase small interfering RNA (siRNA) library (691 kinases) was screened against the TNBC cell line SUM149 for growth inhibition. Selected siRNAs were then tested on four different breast cancer cell lines to confirm the spectrum of activity. Their effect on the CD44high subpopulation and sorted CD44high/CD24-/low cells of SUM149 also was studied. Further studies were focused on polo-like kinase 1 (PLK1), including its expression in breast cancer cell lines, effect on the CD44high/CD24-/low TIC subpopulation, growth inhibition, mammosphere formation, and apoptosis, as well as the activity of the PLK1 inhibitor, BI 2536.

Results

Of the 85 kinases identified in the screen, 28 of them were further silenced by siRNAs on MDA-MB-231 (TNBC), BT474-M1 (ER+/HER2+, a metastatic variant), and HR5 (ER+/HER2+, a trastuzumab-resistant model) cells and showed a broad spectrum of growth inhibition. Importantly, 12 of 28 kinases also reduced the CD44high subpopulation compared with control in SUM149. Further tests of these 12 kinases directly on a sorted CD44high/CD24-/low TIC subpopulation of SUM149 cells confirmed their effect. Blocking PLK1 had the greatest growth inhibition on breast cancer cells and TICs by about 80% to 90% after 72 hours. PLK1 was universally expressed in breast cancer cell lines, representing all of the breast cancer subtypes, and was positively correlated to CD44. The PLK1 inhibitor BI 2536 showed similar effects on growth, mammosphere formation, and apoptosis as did PLK1 siRNAs. Finally, whereas paclitaxel, doxorubicin, and 5-fluorouracil enriched the CD44high/CD24-/low population compared with control in SUM149, subsequent treatment with BI 2536 killed the emergent population, suggesting that it could potentially be used to prevent relapse.

Conclusion

Inhibiting PLK1 with siRNA or BI 2536 blocked growth of TNBCs including the CD44high/CD24-/low TIC subpopulation and mammosphere formation. Thus, PLK1 could be a potential therapeutic target for the treatment of TNBC as well as other subtypes of breast cancer.
Appendix
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Literature
1.
Perou CM, Sorlie T, Eisen MB, van de Rijn M, Jeffrey SS, Rees CA, Pollack JR, Ross DT, Johnsen H, Akslen LA, Fluge O, Pergamenchikov A, Williams C, Zhu SX, Lønning PE, Børreson-Dale AL, Brown PO, Botstein D: Molecular portraits of human breast tumours. Nature. 2000, 406: 747-752. 10.1038/35021093.CrossRefPubMed
2.
Sørlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H, Hastie T, Eisen MB, van de Rijn M, Jeffrey SS, Thorsen T, Quist H, Matese JC, Brown PO, Botstein D, Eystein Lønning P, Børresen-Dale AL: Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci USA. 2001, 98: 10869-10874. 10.1073/pnas.191367098.CrossRefPubMedPubMedCentral
3.
Dent R, Trudeau M, Pritchard KL, Hanna WM, Kahn HK, Sawka CA, Lickley LA, Rawlinson E, Sun P, Narod SA: Triple-negative breast cancer: clinical features and patterns of recurrence. Clin Cancer Res. 2007, 13: 4429-4434. 10.1158/1078-0432.CCR-06-3045.CrossRefPubMed
4.
5.
de Ruijter TC, Veeck J, de Hoon JPJ, van Engeland M, Tjan-Heijnen VC: Characteristics of triple-negative breast cancer. J Cancer Res Clin Oncol. 2011, 137: 183-192. 10.1007/s00432-010-0957-x.CrossRefPubMed
6.
Fong PC, Boss DS, Yap TA, Tutt A, Wu P, Mergui-Roelvink M, Mortimer P, Swasiland H, Lau A, O'Connor MJ, Ashworth A, Carmichael J, Kaye SB, Schellens JH, de Bono JS: Inhibition of poly(ADP-ribosa) polymerase in tumors from BRCA mutation carriers. N Engl J Med. 2009, 361: 123-134. 10.1056/NEJMoa0900212.CrossRefPubMed
7.
Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D, Lopez E, Kyle S, Meuth M, Curtin NJ, Helleday T: Specific killing of BRCA2-deficient tumours with inhibitors of poly (ADP-ribose) polymerase. Nature. 2005, 434: 913-917. 10.1038/nature03443. Erratum, Nature 2007, 447:346CrossRefPubMed
8.
Evans DG, Howell A, Ward D, Lalloo F, Jones JL, Eccles DM: Prevalence of BRCA1 and BRCA2 mutations in triple negative breast cancer. J Med Genet. 2011, doi:10.1136/jmedgenet-2011-100006
9.
Santana-Davila R, Perez EA: Treatment options for patients with triple-negative breast cancer. J Hematol Oncol. 2010, 3: 42-52. 10.1186/1756-8722-3-42.CrossRefPubMedPubMedCentral
10.
11.
Charafe-Jauffret E, Ginestier C, Birnbaum D: Breast cancer stem cells: tools and models to relay on. BMC Cancer. 2009, 9: 202-211. 10.1186/1471-2407-9-202.CrossRefPubMedPubMedCentral
12.
Zhou BS, Zhang H, Damelin M, Geles KG, Grindley JC, Dirks PB: Tumour-initiating cells: challenges and opportunities for anticancer drug discovery. Nature Rev Drug Discovery. 2009, 8: 806-823. 10.1038/nrd2137.CrossRef
13.
Sajithlal GB, Rothermund K, Zhang F, Dabbs DJ, Latimer JJ, Grant SG, Prochownik EV: Permanently blocked stem cells derived from breast cancer cell lines. Stem Cells. 2010, 28: 1008-1018. 10.1002/stem.424.CrossRefPubMedPubMedCentral
14.
To K, Zhao Y, Jiang H, Hu K, Wang M, Wu J, Lee C, Yokom DW, Stratford AL, Klinge U, Mertens PR, Chen CS, Bally M, Yapp D, Dunn SE: The phosphoinositide-dependent kinase-1 inhibitor, OSU-03012, prevents Y-box binding protein-1 (YB-1) from inducing epidermal growth factor receptor (EGFR). Mol Pharmacol. 2007, 72: 641-652. 10.1124/mol.107.036111.CrossRefPubMed
15.
Fillmore CM, Kuperwasser C: Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy. Breast Cancer Res. 2008, 10: R25-10.1186/bcr1982.CrossRefPubMedPubMedCentral
16.
Smith KM, Datti A, Fujitani M, Grinshtein N, Zhang L, Morozova O, Blakely KM, Rotenberg SA, Hansford LM, Miller FD, Yeger H, Irwin MS, Moffat J, Marra MA, Baruchel S, Wrana JL, Kaplan DR: Selective targeting of neuroblastoma tumour-initiating cells by compounds identified in stem cell-based small molecule screens. EMBO Mol Med. 2010, 2: 371-384. 10.1002/emmm.201000093.CrossRefPubMedPubMedCentral
17.
Lee HE, Kim JH, Kim YJ, Choi SY, Kim SW, Kang E, Chung IY, Kim IA, Kim EJ, Choi Y, Ryu HS, Park SY: An increase in cancer stem cell population after primary systemic therapy is a poor prognostic factor in breast cancer. Br J Cancer. 2011, 104: 1730-1738. 10.1038/bjc.2011.159.CrossRefPubMedPubMedCentral
18.
MacKeigan JP, Murphy LO, Blenis J: Sensitized RNAi screen of human kinases and phosphatases identifies new regulators of apoptosis and chemoresistance. Nature Cell Biol. 2005, 7: 591-600. 10.1038/ncb1258.CrossRefPubMed
19.
Hu K, Lee C, Qiu D, Fotovati A, Davies A, Abu-Ali S, Wai D, Lawlor ER, Triche TJ, Pallen CJ, Dunn SE: Small interfering RNA library screen of human kinases and phosphatases identifies polo-like kinase 1 as a promising new target for the treatment of pediatric rhabdomyosarcomas. Mol Cancer Ther. 2009, 8: 3024-3034. 10.1158/1535-7163.MCT-09-0365.CrossRefPubMedPubMedCentral
20.
Bauer JA, Ye F, Marshall CB, Lehmann BD, Pendleton CS, Shyr Y, Arteaga CL, Pietenpol JA: RNA interference (RNAi) screen approach identifies agents that enhance paclitaxel activity in breast cancer cells. Breast Cancer Res. 2010, 12: R41-10.1186/bcr2595.CrossRefPubMedPubMedCentral
21.
Cohen P: Protein kinases: the major drug targets of the twenty-first century?. Nature Rev Drug Discov. 2002, 1: 309-315. 10.1038/nrd773.CrossRef
22.
23.
Speers C, Tsimelzon A, Sexton K, Herrick AM, Gutierrez C, Culhane A, Quackenbush J, Hilsenbeck S, Chang J, Brown P: Identification of novel kinase targets for the treatment of estrogen receptor-negative breast cancer. Clin Cancer Res. 2009, 15: 6327-6340. 10.1158/1078-0432.CCR-09-1107.CrossRefPubMedPubMedCentral
24.
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.CrossRefPubMedPubMedCentral
25.
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, Dewhirst MW, Bigner DD, Rich JN: Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006, 444: 756-760. 10.1038/nature05236.CrossRefPubMed
26.
Grinshtein N, Datti A, Fujitani M, Uehling D, Prakesch M, Isaac M, Irwin MS, Wrana JL, Al-Awar R, Kaplan DR: Small molecule kinase inhibitor screen identifies polo-like kinase 1 as a target for neuroblastoma tumor-initiating cells. Cancer Res. 2011, 71: 1385-1395. 10.1158/0008-5472.CAN-10-2484.CrossRefPubMed
27.
Prochownik EV, Lazo JS: Small molecule inhibitors of cancer stem cells. US Patent. 2010, 20100298352-
28.
Steegmaier M, Hoffmann M, Baum A, Lénárt P, Petronczki M, Krššák M, Gürtler U, Garin-Chesa P, Lieb S, Quant J, Grauert M, Adolf GR, Kraut N, Peters JM, Wolfgang J, Rettig WJ: BI 2536, a potent and selective inhibitor of polo-like kinase 1, inhibits tumor growth in vivo. Curr Biol. 2007, 17: 316-322. 10.1016/j.cub.2006.12.037.CrossRefPubMed
29.
Wu J, Lee C, Yokom D, Jiang H, Cheang MCU, Yorida E, Turbin D, Berquin IM, Mertens P, 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.CrossRefPubMed
30.
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: YB-1 induces 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.CrossRefPubMedPubMedCentral
31.
Abraham BK, Fritz P, McClellan M, Hauptvogel P, Athelogou M, Brauch H: Prevalence of CD44+/CD24-/low cells in breast cancer may not be associated with clinical outcome but may favor distant metastasis. Clin Cancer Res. 2005, 11: 1154-1159.PubMed
32.
Spankuch B, Kurunci-Csacsko E, Kaufmann M, Strebhardt K: Rational combinations of siRNAs targeting plk1 with breast cancer drugs. Oncogene. 2007, 26: 5793-5807. 10.1038/sj.onc.1210355.CrossRefPubMed
33.
Lorns E, Lord CJ, Turner N, Ashworth A: Utilizing RNA interference to enhance cancer drug discovery. Nature Rev. 2007, 6: 556-568. 10.1038/nrd2355.
34.
Signoretti S, Di Marcotullio L, Richardson A, Ramaswamy S, Isaac B, Rue M, Monti F, Loda M, Pagano M: Oncogenic role of the ubiquitin ligase subunit Skp2 in human breast cancer. J Clin Invest. 2002, 110: 633-641.CrossRefPubMedPubMedCentral
35.
Sonoda H, Inoue H, Ogawa K, Utsunomiya T, Masuda TA, Mori M: Significance of skp2 expression in primary breast cancer. Clin Cancer Res. 2006, 12: 1215-1220. 10.1158/1078-0432.CCR-05-1709.CrossRefPubMed
36.
Davidovich S, Ben-Izhak O, Shapira M, Futerman B, Hershko DD: Over-expression of Skp2 is associated with resistance to preoperative doxorubicin-based chemotherapy in primary breast cancer. Breast Cancer Res. 2008, 10: R63-10.1186/bcr2122.CrossRefPubMedPubMedCentral
37.
de Castro PI, de Carcer G, Montoya G, Malumbres M: Emerging cancer therapeutic opportunities by inhibiting mitotic kinases. Curr Opin Pharmacol. 2008, 8: 375-383. 10.1016/j.coph.2008.06.013.CrossRef
38.
39.
Casarsa C, Oriana S, Coradini D: The controversial clinicobiological role of breast cancer stem cells. J Oncol. 2008, 2008: 492643-///CrossRefPubMed
40.
Strebhardt K, Ullrich A: Targeting polo-like kinase 1 for cancer therapy. Nature Rev Cancer. 2006, 6: 321-330. 10.1038/nrc1841.CrossRef
41.
de Castro PI, de Carcer G, Malumbres M: A census of mitotic cancer genes: new insight into tumor cell biology and cancer therapy. Carcinogenesis. 2007, 28: 899-912.CrossRef
42.
Yuan J, Hörlin A, Hock B, Stutte HJ, Rübsamen-Waigmann H, Strebhardt K: Polo-like kinase, a novel marker for cellular proliferation. Am J Pathol. 1997, 150: 1165-1172.PubMedPubMedCentral
43.
Wolf G, Elez R, Doermer A, Holtrich U, Ackermann H, Stutte HJ, Altmannsberger HM, Rübsamen-Waigmann H, Strebhardt K: Prognostic significance of polo-like kinase (PLK) expression in non-small cell lung cancer. Oncogene. 1997, 14: 543-549. 10.1038/sj.onc.1200862.CrossRefPubMed
44.
Kneisel L, Strebhardt K, Bernd A, Wolter M, Binder A, Kaufmann R: Expression of polo-like kinase (PLK1) in thin melanomas: a novel marker of metastatic disease. J Cutan Pathol. 2002, 29: 354-358. 10.1034/j.1600-0560.2002.290605.x.CrossRefPubMed
45.
Ahr A, Holtrich U, Solbach C, Schar A, Strebhardt K, Karn T, Kaufmann M: Molecular classification of breast cancer patients by gene expression profiling. J Pathol. 2001, 195: 312-320. 10.1002/path.955.CrossRefPubMed
46.
Ahr A, Karn T, Solbach C, Seiter T, Strebhardt K, Holtrich U, Kaufmann M: Identification of high risk breast-cancer patients by gene expression profiling. Lancet. 2002, 359: 131-132. 10.1016/S0140-6736(02)07337-3.CrossRefPubMed
47.
Liu X, Lei M, Erikson RL: Normal cells, but not cancer cells, survive severe plk1 depletion. Mol Cell Biol. 2006, 26: 2093-2108. 10.1128/MCB.26.6.2093-2108.2006.CrossRefPubMedPubMedCentral
48.
Degenhardt Y, Lampkin T: Targeting polo-like kinase in cancer therapy. Clin Cancer Res. 2010, 16: 384-389. 10.1158/1078-0432.CCR-09-1380.CrossRefPubMed
49.
Schöffski P, Blay JY, De Greve J, Brain E, Machiels JP, Soria JC, Sleijfer S, Wolter P, Ray-Coquard I, Fontaine C, Munzert G, Fritsch H, Hanft G, Aerts C, Rapion J, Allgeier A, Bogaerts J, Lacombe D: Multicentric parallel phase II trial of the polo-like kinase 1 inhibitor BI 2536 in patients with advanced head and neck cancer, breast cancer, ovarian cancer, soft tissue sarcoma and melanoma: The first protocol of the European Organization for Research and Treatment of Cancer (EORTC) Network Of Core Institutes (NOCI). Eur J Cancer. 2010, 46: 2206-2215. 10.1016/j.ejca.2010.03.039.CrossRefPubMed
50.
Harrison MR, Holen KD, Liu G: Beyond taxanes: a review of novel agents that target mitotic tubulin and microtubules, kinases, and kinesins. Clin Adv Hematol Oncol. 2009, 7: 54-64.PubMedPubMedCentral
51.
Chopra P, Sethi G, Dastidar SG, Ray A: Polo-like kinase inhibitors: an emerging opportunity for cancer therapeutics. Expert Opin Investig Drugs. 2010, 19: 27-43. 10.1517/13543780903483191.CrossRefPubMed
52.
Gleixner KV, Ferenc V, Peter B, Gruze A, Meyer RA, Hadzijusufovic E, Cerny-Reiterer S, Mayerhofer M, Pickl WF, Sillaber C, Valent P: Polo-like kinase 1 (Plk1) as a novel drug target in chronic myeloid leukemia: overriding imatinib resistance with the Plk1 inhibitor BI 2536. Cancer Res. 2010, 70: 1513-1523. 10.1158/0008-5472.CAN-09-2181.CrossRefPubMed
53.
Bevan P, Mala C: The Role of uPA and uPA inhibitors in breast cancer. Breast Care (Basel). 2008, 3 (Suppl 2): 1-2.CrossRef
54.
Goldstein LJ: Experience in phase I trials and an upcoming phase II study with uPA inhibitors in metastatic breast cancer. Breast Care (Basel). 2008, 3 (Suppl 2): 25-28.
55.
Sheridan C, Kishimoto H, Fuchs RK, Mehrotra S, Bhat-Nakshatri P, Turner CH, Goulet R, Badve S, Nakshatri H: CD44+/CD24- breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasis. Breast Cancer Res. 2006, 8: R59-10.1186/bcr1610.CrossRefPubMedPubMedCentral
56.
Balic M, Lin H, Young L, Hawes D, Giuliano A, McNamara G, Datar RH, Cote RJ: Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype. Clin Cancer Res. 2006, 12: 5615-5621. 10.1158/1078-0432.CCR-06-0169.CrossRefPubMed
Metadata
Title
Small interfering RNA library screen identified polo-like kinase-1 (PLK1) as a potential therapeutic target for breast cancer that uniquely eliminates tumor-initiating cells
Authors
Kaiji Hu
Jennifer H Law
Abbas Fotovati
Sandra E Dunn
Publication date
01-02-2012
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 1/2013
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3107

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