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

Molecular Cancer Therapeutics - Tập 8 Số 11 - Trang 3024-3035 - 2009
Kaiji Hu1,2, Cathy Lee1,2, Dexin Qiu1,2, Abbas Fotovati1,2, Alastair Davies1,2, Samah Abu‐Ali1,2, Daniel Wai1,2, Elizabeth R. Lawlor1,2, Timothy J. Triche1,2, Catherine J. Pallen1,2, Sandra E. Dunn1,2
11Laboratory for Oncogenomic Research, Departments of Pediatrics, Experimental Medicine, and Medical Genetics, and 2Cell Phosphosignaling Laboratory, Departments of Pediatrics, Pathology and Laboratory Medicine, and Experimental Medicine, Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia, Canada; and 3Department of Pathology, Keck School of Medicine of the University of Southern California, Los Angeles, California
2Department of Pathology, Keck School of Medicine of the University of Southern California, Los Angeles, California

Tóm tắt

Abstract Rhabdomyosarcoma, consisting of alveolar (aRMS) and embryonal (eRMS) subtypes, is the most common type of sarcoma in children. Currently, there are no targeted drug therapies available for rhabdomyosarcoma. In searching for new molecular therapeutic targets, we carried out genome-wide small interfering RNA (siRNA) library screens targeting human phosphatases (n = 206) and kinases (n = 691) initially against an aRMS cell line, RH30. Sixteen phosphatases and 50 kinases were identified based on growth inhibition after 72 hours. Inhibiting polo-like kinase 1 (PLK1) had the most remarkable impact on growth inhibition (∼80%) and apoptosis on all three rhabdomyosarcoma cell lines tested, namely, RH30, CW9019 (aRMS), and RD (eRMS), whereas there was no effect on normal muscle cells. The loss of PLK1 expression and subsequent growth inhibition correlated with decreased p-CDC25C and Cyclin B1. Increased expression of WEE 1 was also noted. The induction of apoptosis after PLK1 silencing was confirmed by increased p-H2AX, propidium iodide uptake, and chromatin condensation, as well as caspase-3 and poly(ADP-ribose) polymerase cleavage. Pediatric Ewing's sarcoma (TC-32), neuroblastoma (IMR32 and KCNR), and glioblastoma (SF188) models were also highly sensitive to PLK1 inhibition. Finally, based on cDNA microarray analyses, PLK1 mRNA was overexpressed (>1.5 fold) in 10 of 10 rhabdomyosarcoma cell lines and in 47% and 51% of primary aRMS (17 of 36 samples) and eRMS (21 of 41 samples) tumors, respectively, compared with normal muscles. Similarly, pediatric Ewing's sarcoma, neuroblastoma, and osteosarcoma tumors expressed high PLK1. We conclude that PLK1 could be a promising therapeutic target for the treatment of a wide range of pediatric solid tumors including rhabdomyosarcoma. [Mol Cancer Ther 2009;8(11):3024–35]

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Tài liệu tham khảo

Paulino, 2008, Rhabdomyosarcoma, Curr Probl Cancer, 32, 7, 10.1016/j.currproblcancer.2007.11.001

Merlino, 1999, Rhabdomyosarcoma-working out the pathways, Oncogene, 18, 5340, 10.1038/sj.onc.1203038

Pappo, 1995, Biology and therapy of pediatric rhabdomyosarcoma, J Clin Oncol, 13, 2123, 10.1200/JCO.1995.13.8.2123

Davicioni, 2006, Identification of a PAX-FKHR gene expression signature that defines molecular classes and determines the prognosis of alveolar rhabdomyosarcomas, Cancer Res, 66, 6936, 10.1158/0008-5472.CAN-05-4578

Kurmasheva, 2007, Pediatric oncology, Curr Opin Chem Biol, 11, 424, 10.1016/j.cbpa.2007.05.037

Blandford, 2006, Rhabdomyosarcomas utilize developmental, myogenic growth factors for disease advantage: a report from the Children's Oncology Group, Pediatr Blood Cancer, 46, 329, 10.1002/pbc.20466

Melcon, 2007, Molecular biology of rhabdomyosarcoma, Clin Transl Oncol, 9, 415, 10.1007/s12094-007-0079-3

Scrable, 1987, Chromosomal localization of the human rhabdomyosarcoma locus by mitotic recombination mapping, Nature, 329, 645, 10.1038/329645a0

Barr, 1997, Molecular genetics and pathogenesis of rhabdomyosarcoma, J Pediatr Hematol Oncol, 19, 483, 10.1097/00043426-199711000-00001

Bennicelli, 1995, Wild-type PAX3 protein and the PAX3-FKHR fusion protein of alveolar rhabdomyosarcoma contain potent, structurally distinct transcriptional activation domains, Oncogene, 11, 119

Fredericks, 1995, The PAX3-FKHR fusion protein created by the t(2;13) translocation in alveolar rhabdomyosarcoma is a more potent transcriptional activator than PAX3, Mol Cell Biol, 15, 1522, 10.1128/MCB.15.3.1522

Khan, 2001, Classification and diagnostic prediction of cancers using gene expression profiling and artificial neural networks, Nat Med, 7, 673, 10.1038/89044

Baer, 2004, Profiling and functional annotation of mRNA gene expression in pediatric rhabdomyosarcoma and Ewing's sarcoma, Int J Cancer, 110, 687, 10.1002/ijc.20171

Wachtel, 2004, Gene expression signatures identify rhabdomyosarcoma subtypes and detect a novel t(2;2)(q35;p23) translocation fusing PAX3 to NCOA1, Cancer Res, 64, 5539, 10.1158/0008-5472.CAN-04-0844

De Pittà, 2006, Gene expression profiling identifies potential relevant genes in alveolar rhabdomyosarcoma pathogenesis and discriminates PAX3-FKHR positive and negative tumors, Int J Cancer, 118, 2772, 10.1002/ijc.21698

Schaaf, 2005, Full transcriptome analysis of rhabdomyosarcoma, normal, and fetal skeletal muscle: statistical comparison of multiple SAGE libraries, FASEB J, 19, 404, 10.1096/fj.04-2104fje

Goldstein, 2006, Novel genes implicated in embryonal, alveolar, and pleomorphic rhabdomyosarcoma: a cytogenetic and molecular analysis of primary tumors, Neoplasia, 8, 332, 10.1593/neo.05829

Ebauer, 2007, Comparative expression profiling identifies an in vivo target gene signature with TFAP2B as a mediator of the survival function of PAX3/FKHR, Oncogene, 26, 7267, 10.1038/sj.onc.1210525

Romualdi, 2006, Defining the gene expression signature of rhabdomyosarcoma by meta-analysis, BMC Genomics, 7, 287, 10.1186/1471-2164-7-287

Ren, 2008, Mouse mesenchymal stem cells expressing PAX-FKHR form alveolar rhabdomyosarcomas by cooperating with secondary mutations, Cancer Res, 68, 6587, 10.1158/0008-5472.CAN-08-0859

MacKeigan, 2005, Sensitized RNAi screen of human kinases and phosphatases identifies new regulators of apoptosis and chemoresistance, Nat Cell Biol, 7, 591, 10.1038/ncb1258

Saab, 2006, Pharmacologic inhibition of cyclin-dependent kinase 4/6 activity arrests proliferation in myoblasts and rhabdomyosarcoma-derived cells, Mol Cancer Ther, 5, 1299, 10.1158/1535-7163.MCT-05-0383

Amstutz, 2008, Phosphorylation regulates transcriptional activity of PAX3/FKHR and reveals novel therapeutic possibilities, Cancer Res, 68, 3767, 10.1158/0008-5472.CAN-07-2447

Cao, 2008, Addiction of elevated insulin-like growth factor I receptor and initial modulation of the AKT pathway define the responsiveness of rhabdomyosarcoma to the targeting antibody, Cancer Res, 68, 8039, 10.1158/0008-5472.CAN-08-1712

Cen, 2007, Phosphorylation profiles of protein kinases in alveolar and embryonal rhabdomyosarcoma, Mod Pathol, 20, 936, 10.1038/modpathol.3800834

To, 2007, 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, 72, 641, 10.1124/mol.107.036111

Lee, 2008, Targeting YB-1 in HER-2 overexpressing breast cancer cells induces apoptosis via the mTOR/STAT3 pathway and suppresses tumor growth in mice, Cancer Res, 68, 8661, 10.1158/0008-5472.CAN-08-1082

Newton, 1995, Classification of rhabdomyosarcomas and related sarcomas. Pathologic aspects and proposal for a new classification - an intergroup rhabdomyosarcoma study, Cancer, 76, 1073, 10.1002/1097-0142(19950915)76:6<1073::AID-CNCR2820760624>3.0.CO;2-L

Santos, 2007, Urokinase-type Plasminogen Activator (uPA) is Inhibited with QLT0267 a Small Molecule Targeting Integrin-linked Kinase (ILK), Translational Oncogenomics, 2, 67

Davicioni, 2009, Molecular classification of rhabdomyosarcoma-genotypic and phenotypic determinants of diagnosis: a report from the Children's Oncology Group, Am J Pathol, 174, 550, 10.2353/ajpath.2009.080631

Taniguchi, 2008, PDGFR-A is a therapeutic target in alveolar rhabdomyosarcoma, Oncogene, 27, 6550, 10.1038/onc.2008.255

Wan, 2006, CCI-779 inhibits rhabdomyosarcoma xenograft growth by an antiangiogenic mechanism linked to the targeting of mTOR/Hif-1α/VEGF signaling, Neoplasia, 8, 394, 10.1593/neo.05820

Chen, 2007, Signal transducer and activator of transcription 3 is involved in cell growth and survival of human rhabdomyosarcoma and osteosarcoma cells, BMC Cancer, 7, 111, 10.1186/1471-2407-7-111

Petricoin, 2007, Phosphoprotein pathway mapping: Akt/mammalian target of rapamycin activation is negatively associated with childhood rhabdomyosarcoma survival, Cancer Res, 67, 3431, 10.1158/0008-5472.CAN-06-1344

Elia, 2008, PI3K/Akt pathway activation attenuates the cytotoxic effect of methyl jasmonate toward sarcoma cells, Neoplasia, 10, 1303, 10.1593/neo.08636

Strebhardt, 2006, Targeting polo-like kinase 1 for cancer therapy, Nat Rev Cancer, 6, 321, 10.1038/nrc1841

Perez de Castro, 2007, A census of mitotic cancer genes: new insight into tumor cell biology and cancer therapy, Carcionogenesis, 28, 899, 10.1093/carcin/bgm019

Perez de Castro, 2008, Emerging cancer therapeutic opportunities by inhibiting mitotic kinases, Curr Opin Pharmacol, 8, 375, 10.1016/j.coph.2008.06.013

Reindl, 2008, Inhibition of polo-like kinase 1 by blocking polo-box domain-dependent protein-protein interactions, Chem Biol, 15, 459, 10.1016/j.chembiol.2008.03.013

Liu, 2003, Polo-like kinase (Plk)1 depletion induces apoptosis in cancer cells, Proc Natl Acad Sci U S A, 100, 5789, 10.1073/pnas.1031523100

Liu, 2006, Normal cells, but not cancer cells, survive severe Plk1 depletion, Mol Cell Biol, 26, 2093, 10.1128/MCB.26.6.2093-2108.2006

Toyoshima-Morimoto, 2002, Plk1 promotes nuclear translocation of human Cdc25C during prophase, EMBO Rep, 3, 341, 10.1093/embo-reports/kvf069

Toyoshima-Morimoto, 2001, Polo-like kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase, Nature, 410, 215, 10.1038/35065617

Reagan-Shaw, 2005, Silencing of polo-like kinase (Plk) 1 via siRNA causes induction of apoptosis and impairment of mitosis machinery in human prostate cancer cells: implications for the treatment of prostate cancer, FASEB J, 19, 611, 10.1096/fj.04-2910fje

Bu, 2008, Silencing of polo-like (Plk) 1 via siRNA causes inhibition of growth and induction of apoptosis in human esophageal cancer cells, Oncology, 74, 198, 10.1159/000151367

Feng, 2009, Overexpression of PLK1 is associated with poor survival by inhibiting apoptosis via enhancement of survivin level in esophageal squamous cell carcinoma, Int J Cancer, 124, 578, 10.1002/ijc.23990

Olmos, 2008, Targeting polo-like kinase: learning too little and too late?, J Clinical Oncol, 26, 5497, 10.1200/JCO.2008.18.6585

Taylor, 2000, p53 mutation and MDM2 amplification frequency in pediatric rhabdomyosarcoma tumors and cell lines, Med Pediatr Oncol, 35, 96, 10.1002/1096-911X(200008)35:2<96::AID-MPO2>3.0.CO;2-Z

Caldas, 2006, Survivin-directed RNA interference cocktail is a potent suppressor of tumour growth in vivo, J Med Genet, 43, 119, 10.1136/jmg.2005.034686

Ikezoe, 2009 :23, A novel treatment strategy targeting polo-like kinase 1 in hematological malignancies, Leukemia, 1564, 10.1038/leu.2009.94

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Molecular Cancer Therapeutics

Tập 8 Số 11

3024-3035

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