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BMC Cancer
Published in: BMC Cancer 1/2010

Open Access 01-12-2010 | Research article

17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells

Authors: Panagiotis K Karkoulis, Dimitrios J Stravopodis, Lukas H Margaritis, Gerassimos E Voutsinas

Published in: BMC Cancer | Issue 1/2010

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Abstract

Background

17-Allylamino-17-demethoxygeldanamycin (17-AAG), a benzoquinone ansamycin antibiotic, specifically targets heat shock protein 90 (Hsp90) and interferes with its function as a molecular chaperone that maintains the structural and functional integrity of various protein clients involved in cellular signaling. In this study, we have investigated the effect of 17-AAG on the regulation of Hsp90-dependent signaling pathways directly implicated in cell cycle progression, survival and motility of human urinary bladder cancer cell lines.

Methods

We have used MTT-based assays, FACS analysis, Western blotting, semi-quantitative RT-PCR, immunocytochemistry and scratch-wound assay in RT4, RT112 and T24 human urinary bladder cancer cell lines.

Results

We have demonstrated that, upon 17-AAG treatment, bladder cancer cells are arrested in the G1 phase of the cell cycle and eventually undergo apoptotic cell death in a dose-dependent manner. Furthermore, 17-AAG administration was shown to induce a pronounced downregulation of multiple Hsp90 protein clients and other downstream effectors, such as IGF-IR, Akt, IKK-α, IKK-β, FOXO1, ERK1/2 and c-Met, resulting in sequestration-mediated inactivation of NF-κB, reduced cell proliferation and decline of cell motility.

Conclusions

In total, we have clearly evinced a dose-dependent and cell type-specific effect of 17-AAG on cell cycle progression, survival and motility of human bladder cancer cells, due to downregulation of multiple Hsp90 clients and subsequent disruption of signaling integrity.
Appendix
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Literature
1.
2.
Wu XR: Urothelial tumorigenesis: a tale of divergent pathways. Nat Rev Cancer. 2005, 5 (9): 713-725. 10.1038/nrc1697.CrossRefPubMed
3.
Pashos CL, Botteman MF, Laskin BL, Redaelli A: Bladder cancer: epidemiology, diagnosis, and management. Cancer Pract. 2002, 10 (6): 311-322. 10.1046/j.1523-5394.2002.106011.x.CrossRefPubMed
4.
Kaufman DS, Shipley WU, Feldman AS: Bladder cancer. Lancet. 2009, 374 (9685): 239-249. 10.1016/S0140-6736(09)60491-8.CrossRefPubMed
5.
Mitra AP, Datar RH, Cote RJ: Molecular staging of bladder cancer. BJU Int. 2005, 96 (1): 7-12. 10.1111/j.1464-410X.2005.05557.x.CrossRefPubMed
6.
Stravopodis DJ, Margaritis LH, Voutsinas GE: Drug-mediated targeted disruption of multiple protein activities through functional inhibition of the Hsp90 chaperone complex. Curr Med Chem. 2007, 14 (29): 3122-3138. 10.2174/092986707782793925.CrossRefPubMed
7.
Whitesell L, Lindquist SL: HSP90 and the chaperoning of cancer. Nat Rev Cancer. 2005, 5 (10): 761-772. 10.1038/nrc1716.CrossRefPubMed
8.
Picard D: Heat-shock protein 90, a chaperone for folding and regulation. Cell Mol Life Sci. 2002, 59 (10): 1640-1648. 10.1007/PL00012491.CrossRefPubMed
9.
Pearl LH, Prodromou C: Structure and mechanism of the Hsp90 molecular chaperone machinery. Annu Rev Biochem. 2006, 75: 271-294. 10.1146/annurev.biochem.75.103004.142738.CrossRefPubMed
10.
Maloney A, Workman P: HSP90 as a new therapeutic target for cancer therapy: the story unfolds. Expert Opin Biol Ther. 2002, 2 (1): 3-24. 10.1517/14712598.2.1.3.CrossRefPubMed
11.
Neckers L, Ivy SP: Heat shock protein 90. Curr Opin Oncol. 2003, 15 (6): 419-424. 10.1097/00001622-200311000-00003.CrossRefPubMed
12.
Powers MV, Workman P: Targeting of multiple signalling pathways by heat shock protein 90 molecular chaperone inhibitors. Endocr Relat Cancer. 2006, 13 (Suppl 1): S125-135. 10.1677/erc.1.01324.CrossRefPubMed
13.
Workman P, Burrows F, Neckers L, Rosen N: Drugging the cancer chaperone HSP90: combinatorial therapeutic exploitation of oncogene addiction and tumor stress. Ann N Y Acad Sci. 2007, 1113: 202-216. 10.1196/annals.1391.012.CrossRefPubMed
14.
Stravopodis DJ, Karkoulis PK, Konstantakou EG, Melachroinou S, Lampidonis AD, Anastasiou D, Kachrilas S, Messini-Nikolaki N, Papassideri IS, Aravantinos G, et al: Grade-dependent effects on cell cycle progression and apoptosis in response to doxorubicin in human bladder cancer cell lines. Int J Oncol. 2009, 34 (1): 137-160.PubMed
15.
Konstantakou EG, Voutsinas GE, Karkoulis PK, Aravantinos G, Margaritis LH, Stravopodis DJ: Human bladder cancer cells undergo cisplatin-induced apoptosis that is associated with p53-dependent and p53-independent responses. Int J Oncol. 2009, 35 (2): 401-416.PubMed
16.
Morales JL, Perdew GH: Carboxyl terminus of hsc70-interacting protein (CHIP) can remodel mature aryl hydrocarbon receptor (AhR) complexes and mediate ubiquitination of both the AhR and the 90 kDa heat-shock protein (hsp90) in vitro. Biochemistry. 2007, 46 (2): 610-621. 10.1021/bi062165b.CrossRefPubMedPubMedCentral
17.
Jiang J, Ballinger CA, Wu Y, Dai Q, Cyr DM, Hohfeld J, Patterson C: CHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation. J Biol Chem. 2001, 276 (46): 42938-42944. 10.1074/jbc.M101968200.CrossRefPubMed
18.
Kundrat L, Regan L: Identification of residues on Hsp70 and Hsp90 ubiquitinated by the cochaperone CHIP. J Mol Biol. 395 (3): 587-594. 10.1016/j.jmb.2009.11.017.
19.
Giustiniani J, Daire V, Cantaloube I, Durand G, Pous C, Perdiz D, Baillet A: Tubulin acetylation favors Hsp90 recruitment to microtubules and stimulates the signaling function of the Hsp90 clients Akt/PKB and p53. Cell Signal. 2009, 21 (4): 529-539. 10.1016/j.cellsig.2008.12.004.CrossRefPubMed
20.
Okamoto J, Mikami I, Tominaga Y, Kuchenbecker KM, Lin YC, Bravo DT, Clement G, Yagui-Beltran A, Ray MR, Koizumi K, et al: Inhibition of Hsp90 leads to cell cycle arrest and apoptosis in human malignant pleural mesothelioma. J Thorac Oncol. 2008, 3 (10): 1089-1095. 10.1097/JTO.0b013e3181839693.CrossRefPubMedPubMedCentral
21.
Basso AD, Solit DB, Munster PN, Rosen N: Ansamycin antibiotics inhibit Akt activation and cyclin D expression in breast cancer cells that overexpress HER2. Oncogene. 2002, 21 (8): 1159-1166. 10.1038/sj.onc.1205184.CrossRefPubMedPubMedCentral
22.
Burrows F, Zhang H, Kamal A: Hsp90 activation and cell cycle regulation. Cell Cycle. 2004, 3 (12): 1530-1536.CrossRefPubMed
23.
Garcia-Morales P, Carrasco-Garcia E, Ruiz-Rico P, Martinez-Mira R, Menendez-Gutierrez MP, Ferragut JA, Saceda M, Martinez-Lacaci I: Inhibition of Hsp90 function by ansamycins causes downregulation of cdc2 and cdc25c and G(2)/M arrest in glioblastoma cell lines. Oncogene. 2007, 26 (51): 7185-7193. 10.1038/sj.onc.1210534.CrossRefPubMed
24.
Hostein I, Robertson D, DiStefano F, Workman P, Clarke PA: Inhibition of signal transduction by the Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin results in cytostasis and apoptosis. Cancer Res. 2001, 61 (10): 4003-4009.PubMed
25.
Mediavilla-Varela M, Pacheco FJ, Almaguel F, Perez J, Sahakian E, Daniels TR, Leoh LS, Padilla A, Wall NR, Lilly MB, et al: Docetaxel-induced prostate cancer cell death involves concomitant activation of caspase and lysosomal pathways and is attenuated by LEDGF/p75. Mol Cancer. 2009, 8 (1): 68-10.1186/1476-4598-8-68.CrossRefPubMedPubMedCentral
26.
Byun JY, Kim MJ, Yoon CH, Cha H, Yoon G, Lee SJ: Oncogenic Ras signals through activation of both phosphoinositide 3-kinase and Rac1 to induce c-Jun NH2-terminal kinase-mediated, caspase-independent cell death. Mol Cancer Res. 2009, 7 (9): 1534-1542. 10.1158/1541-7786.MCR-08-0542.CrossRefPubMed
27.
Cardillo MR, Sale P, Di Silverio F: Heat shock protein-90, IL-6 and IL-10 in bladder cancer. Anticancer Res. 2000, 20 (6B): 4579-4583.PubMed
28.
Lebret T, Watson RW, Molinie V, O'Neill A, Gabriel C, Fitzpatrick JM, Botto H: Heat shock proteins HSP27, HSP60, HSP70, and HSP90: expression in bladder carcinoma. Cancer. 2003, 98 (5): 970-977. 10.1002/cncr.11594.CrossRefPubMed
29.
Lebret T, Watson RW, Molinie V, Poulain JE, O'Neill A, Fitzpatrick JM, Botto H: HSP90 expression: a new predictive factor for BCG response in stage Ta-T1 grade 3 bladder tumours. Eur Urol. 2007, 51 (1): 161-166. 10.1016/j.eururo.2006.06.006. discussion 166-167CrossRefPubMed
30.
Kamal A, Thao L, Sensintaffar J, Zhang L, Boehm MF, Fritz LC, Burrows FJ: A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors. Nature. 2003, 425 (6956): 407-410. 10.1038/nature01913.CrossRefPubMed
31.
Clarke PA, Hostein I, Banerji U, Stefano FD, Maloney A, Walton M, Judson I, Workman P: Gene expression profiling of human colon cancer cells following inhibition of signal transduction by 17-allylamino-17-demethoxygeldanamycin, an inhibitor of the hsp90 molecular chaperone. Oncogene. 2000, 19 (36): 4125-4133. 10.1038/sj.onc.1203753.CrossRefPubMed
32.
McCollum AK, Teneyck CJ, Sauer BM, Toft DO, Erlichman C: Up-regulation of heat shock protein 27 induces resistance to 17-allylamino-demethoxygeldanamycin through a glutathione-mediated mechanism. Cancer Res. 2006, 66 (22): 10967-10975. 10.1158/0008-5472.CAN-06-1629.CrossRefPubMed
33.
Beck R, Verrax J, Gonze T, Zappone M, Pedrosa RC, Taper H, Feron O, Calderon PB: Hsp90 cleavage by an oxidative stress leads to its client proteins degradation and cancer cell death. Biochem Pharmacol. 2009, 77 (3): 375-383. 10.1016/j.bcp.2008.10.019.CrossRefPubMed
34.
Shen SC, Yang LY, Lin HY, Wu CY, Su TH, Chen YC: Reactive oxygen species-dependent HSP90 protein cleavage participates in arsenical As(+3)- and MMA(+3)-induced apoptosis through inhibition of telomerase activity via JNK activation. Toxicol Appl Pharmacol. 2008, 229 (2): 239-251. 10.1016/j.taap.2008.01.018.CrossRefPubMed
35.
Brader S, Eccles SA: Phosphoinositide 3-kinase signalling pathways in tumor progression, invasion and angiogenesis. Tumori. 2004, 90 (1): 2-8.PubMed
36.
Gazitt Y, Kolaparthi V, Moncada K, Thomas C, Freeman J: Targeted therapy of human osteosarcoma with 17AAG or rapamycin: characterization of induced apoptosis and inhibition of mTOR and Akt/MAPK/Wnt pathways. Int J Oncol. 2009, 34 (2): 551-561.PubMed
37.
Lang SA, Klein D, Moser C, Gaumann A, Glockzin G, Dahlke MH, Dietmaier W, Bolder U, Schlitt HJ, Geissler EK, et al: Inhibition of heat shock protein 90 impairs epidermal growth factor-mediated signaling in gastric cancer cells and reduces tumor growth and vascularization in vivo. Mol Cancer Ther. 2007, 6 (3): 1123-1132. 10.1158/1535-7163.MCT-06-0628.CrossRefPubMed
38.
Hovelmann S, Beckers TL, Schmidt M: Molecular alterations in apoptotic pathways after PKB/Akt-mediated chemoresistance in NCI H460 cells. Br J Cancer. 2004, 90 (12): 2370-2377.PubMedPubMedCentral
39.
Georgakis GV, Li Y, Rassidakis GZ, Martinez-Valdez H, Medeiros LJ, Younes A: Inhibition of heat shock protein 90 function by 17-allylamino-17-demethoxy-geldanamycin in Hodgkin's lymphoma cells down-regulates Akt kinase, dephosphorylates extracellular signal-regulated kinase, and induces cell cycle arrest and cell death. Clin Cancer Res. 2006, 12 (2): 584-590. 10.1158/1078-0432.CCR-05-1194.CrossRefPubMed
40.
Jia W, Yu C, Rahmani M, Krystal G, Sausville EA, Dent P, Grant S: Synergistic antileukemic interactions between 17-AAG and UCN-01 involve interruption of RAF/MEK- and AKT-related pathways. Blood. 2003, 102 (5): 1824-1832. 10.1182/blood-2002-12-3785.CrossRefPubMed
41.
Xie Q, Gao CF, Shinomiya N, Sausville E, Hay R, Gustafson M, Shen Y, Wenkert D, Vande Woude GF: Geldanamycins exquisitely inhibit HGF/SF-mediated tumor cell invasion. Oncogene. 2005, 24 (23): 3697-3707. 10.1038/sj.onc.1208499.CrossRefPubMed
42.
Wang S, Pashtan I, Tsutsumi S, Xu W, Neckers L: Cancer cells harboring MET gene amplification activate alternative signaling pathways to escape MET inhibition but remain sensitive to Hsp90 inhibitors. Cell Cycle. 2009, 8 (13): 2050-2056.CrossRefPubMed
43.
Peruzzi B, Bottaro DP: Targeting the c-Met signaling pathway in cancer. Clin Cancer Res. 2006, 12 (12): 3657-3660. 10.1158/1078-0432.CCR-06-0818.CrossRefPubMed
Metadata
Title
17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells
Authors
Panagiotis K Karkoulis
Dimitrios J Stravopodis
Lukas H Margaritis
Gerassimos E Voutsinas
Publication date
01-12-2010
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2010
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-10-481

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