World Journal of Urology

, Volume 23, Issue 1, pp 38–46 | Cite as

Use of antisense oligonucleotides targeting the cytoprotective gene, clusterin, to enhance androgen- and chemo-sensitivity in prostate cancer

  • Martin GleaveEmail author
  • Hideaki Miyake
Topic Paper


The discovery and targeting of genes mediating androgen-independence may lead to the development of novel therapies that delay progression of hormone refractory prostate cancer (HRPC). Clusterin is a stress-associated cell survival gene that increases after androgen ablation. Here, we review clusterin’s functional role in apoptosis and the use of antisense oligonucleotides (ASOs) against clusterin to enhance apoptosis in prostate cancer models. Immunostaining of tissue microarrays constructed from untreated and post-hormone treated radical prostatectomy specimens confirm that clusterin is highly expressed in virtually all HRPC cells, 80% of prostate cancer cells after neoadjuvant hormone therapy, but is low or absent (<20%) in untreated specimens. Overexpression of clusterin in LNCaP cells confers resistance to both androgen ablation and chemotherapy. Clusterin ASOs reduced clusterin levels in a dose-dependent and sequence-specific manner. Adjuvant treatment with murine clusterin ASOs after castration of mice bearing Shionogi tumors decreased clusterin levels, accelerated apoptotic tumor regression, and significantly delayed the recurrence of androgen-independent tumors. A human clusterin ASO targeting the translation initiation site and incorporating MOE-gapmer backbone (OGX-011) synergistically enhanced the cytotoxic effects of paclitaxel in human xenografts of prostate, renal cell, bladder, and lung cancer. Clusterin, is an anti-apoptosis protein upregulated in an adaptive cell survival manner by androgen ablation and chemotherapy that confers resistance to various cell death triggers. Suppression of clusterin levels using ASOs enhances cell death following treatment with androgen ablation, radiation, and chemotherapy.


Oligonucleotide Clusterin Prostate cancer 


  1. 1.
    Denis L, Murphy GP (1993) Overview of phase III trials on combined androgen treatment in patients with metastatic prostate cancer. Cancer 72:3888–3895Google Scholar
  2. 2.
    Oh WK, Kantoff PW (1998) Management of hormone refractory prostate cancer: current standards and future prospects. J Urol 160:1220–1229Google Scholar
  3. 3.
    Tannock IF, De Wit R, Berry WR et al. for the TAX 327 Investigators (2004) Docetaxel plus prednisone or mitoxantrone plus prednisone for advanced prostate cancer. N Engl J Med 351:1502–1512Google Scholar
  4. 4.
    Petrylak DP, Tangen CM, Hussain MH et al. (2004) Docetaxel and estramustine compared with mitoxantrone and prednisone for advanced refractory prostate cancer. N Engl J Med 351:1513–1520Google Scholar
  5. 5.
    Isaacs JT, Wake N, Coffey DS, Sandberg AA (1982) Genetic instability coupled to clonal selection as a mechanism for progression in prostatic cancer. Cancer Res 42–48:2353Google Scholar
  6. 6.
    Miyake H, Tolcher A, Gleave ME (1999) Antisense Bcl-2 oligodeoxynucleotides delay progression to androgen-independence after castration in the androgen dependent Shionogi tumor model. Cancer Res 59:4030–4034Google Scholar
  7. 7.
    Miyake H, Rennie P, Nelson C, Gleave ME (2000) Testosterone-repressed prostate message-2 (TRPM-2) is an antiapoptotic gene that confers resistance to androgen ablation in prostate cancer xenograft models. Cancer Res 60:170–176Google Scholar
  8. 8.
    Bruchovsky N, Rennie PS, Coldman AJ, Goldenberg SL, To M, Lawson D (1990) Effects of androgen withdrawal on the stem cell composition of the Shionogi carcinoma. Cancer Res 50:2275–2282Google Scholar
  9. 9.
    Miyake H, Nelson C, Rennie P, Gleave ME (2000) Overexpression of insulin-like growth factor binding protein-5 helps accelerate progression to androgen-independence in the human prostate LNCaP tumor model through activation of phosphatidylinositol 3′-kinase pathway. Endocrinology 141:2257–2265CrossRefGoogle Scholar
  10. 10.
    Kiyama S, Morrison K, Zellweger T, Akbari M, Cox ME, Yu D, Miyake H, Gleave ME (2003) Castration-induced increased in insulin-like growth factor-binding protein 2 promotes proliferation of androgen-independent human prostate LNCaP tumors. Cancer Res 63:3575–3584Google Scholar
  11. 11.
    Sato N, Sadar MD, Bruchovsky N, Saatcioglu F, Rennie PS, Sato S, Lange PH, Gleave ME (1997) Androgenic induction of prostate-specific antigen gene is repressed by protein-protein interaction between androgen receptor and AP-1c-Jun in the human prostate cancer cell line LNCaP. J Biol Chem 272:17485–17494Google Scholar
  12. 12.
    Craft N, Shostak Y, Carey M, Sawyers C (1999) A mechanism for hormone-independent prostate cancer through modulation of androgen receptor signaling by the HER-2/neu tyrosine kinase. Nat Med 5:280–285CrossRefGoogle Scholar
  13. 13.
    Sherwood ER, Van Dongen JL, Wood CG, Liao S, Kozlowski JM, Lee C (1998) Epidermal growth factor receptor activation in androgen-independent but not androgen-stimulated growth of human prostatic carcinoma cells. Br J Cancer 77:855–861Google Scholar
  14. 14.
    Abreu-Martin MT, Chari A, Palladino AA, Craft NA, Sawyers CL (1999) Mitogen-activated protein kinase 1 activates androgen receptor-dependent transcription and apoptosis in prostate cancer. Mol Cell Biol 19:5143–5154Google Scholar
  15. 15.
    Chen CD, Welsbie DS, Tran C, Baek SH, Chen R, Vessella R, Rosenfeld MG, Sawyers CL (2004) Molecular determinants of resistance to antiandrogen therapy. Nat Med 10:33–39CrossRefGoogle Scholar
  16. 16.
    Ettinger SL, Sobel R, Whitmore T, Akbari M, Bradley D, Gleave M, Nelson C (2004) Dysregulation of sterol response element binding proteins and downstream effectors in prostate cancer during progression to androgen-independence. Cancer Res 64:2212–21Google Scholar
  17. 17.
    Gimenez-Bonafe, P, Fedoruk MN, WhitmoreTG, Akbari M, Ralph JL, Ettinger S, Gleave ME, Nelson CC (2004) Up-regulation of the transcription factor YB-1 and its correlation with P-glycoprotein expression during prostate cancer tumor progression. Prostate 59:337–349Google Scholar
  18. 18.
    Gleave ME, Tolcher A, Miyake H, Beraldi E, Goldie J (1999) Progression to androgen-independence is delayed by antisense bcl-2 oligodeoxynucleotides after castration in the LNCaP prostate tumor model. Clin Cancer Res 5:2891–2898Google Scholar
  19. 19.
    Paterson R, Gleave M, Jone E, Zubovits J, Goldenberg SL, Sullivan LD (1999) Immunohistochemical analysis of radical prostatectomy specimens after 8 months of neoadjuvant hormone therapy. Mol Urol 3:277–286Google Scholar
  20. 20.
    McDonnell TJ, Troncoso P, Brisby SM, Logothetis CL, Chung LWK, Hsieh JT et al. (1992) Expression of the protooncogene Bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res 52:6940–6944Google Scholar
  21. 21.
    Colombel M, Symmans F, Gil S, O’Toole KM, Choplin D, Benson M et al. (1993) Detection of the apoptosis-suppressing oncoprotein Bcl-2 in hormone-refractory human prostate cancers. Am J Pathol 143:390–400Google Scholar
  22. 22.
    Raffo AJ, Periman H, Chen MW, Streitman JS, Buttyan R (1995) Overexpression of bcl-2 protects prostate cancer cells from apoptosis in vitro and confers resistance to androgen depletion in vivo. Cancer Res 55:4438–4445Google Scholar
  23. 23.
    Jansen B, Schlagbauer-Wadl H, Brown BD, Bryan RN, Van Elsas A, Muller M et al. (1998) bcl-2 antisense therapy chemosensitizes human melanoma in SCID mice. Nat Med 4:232–234Google Scholar
  24. 24.
    Miyake H, Tolcher A, Gleave ME (2000) Antisense bcl-2 oligodeoxynucleotides enhance taxol chemosensitivity and synergistically delays progression to androgen-independence after castration in the androgen dependent Shionogi tumor model. J Natl Cancer Inst 92:34–41CrossRefGoogle Scholar
  25. 25.
    Rocchi P, So A, Kojima S. Beraldi E, Fazli L, Gleave ME (2004) Heat shock protein 27 increases after androgen ablation and plays a cytoprotective role in hormone refractory prostate cancer. Cancer Res 64:6595–6602Google Scholar
  26. 26.
    Nickerson T, Miyake H, Gleave ME, Pollak M (1999) Castration-induced apoptosis of androgen-dependent Shionogi carcinoma is associated with increased expression of genes encoding insulin-like growth factor binding proteins. Cancer Res 59:3392–3395Google Scholar
  27. 27.
    Miyake H, Pollak, Gleave ME (2000) Castration-induced up-regulation of insulin-like growth factor binding protein-5 potentiates insulin-like growth factor-I activity and accelerates progression to androgen independence in prostate cancer models. Cancer Res 60:3058–3064Google Scholar
  28. 28.
    Tenniswood M, Wang Z, Lakins J, Morrissey C, O’Sullivan J, Tang H (1998) Clusterin in the male reproductive tract. J Androl 19:508–516Google Scholar
  29. 29.
    Calero M, Rostagno A, Matsubara E, Zlokovic B, Frangione B, Ghiso J (2000) Apolipoprotein J (clusterin) and Alzheimer’s disease. Microsc Res Tech 50:305–315Google Scholar
  30. 30.
    Rosenberg ME, Silkensen J (1995) Clusterin and the kidney. Exp Nephrol 3: 9–14Google Scholar
  31. 31.
    Steinberg J, Oyasu R, Lang S, Sintich S, Rademaker A, Lee C, Kozlowski JM, Sensibar JA (1997) Intracellular levels of SGP-2 (clusterin) correlate with tumor grade in prostate cancer. Clin Cancer Res 3:1701–1711Google Scholar
  32. 32.
    Cervellera M, Raschella G, Santilli G, Tanno B, Ventura A, Mancini C, Sevignani C, Calabretta B, Sala A (2000) Direct transactivation of the anti-apoptotic gene apolipoprotein J (clusterin) by B-MYB. J Biol Chem 275:21055–21060CrossRefGoogle Scholar
  33. 33.
    Kyprianou N, English HF, Davidson NE, Isaacs JT (1991) Programmed cell death during regression of the MCF-7 human breast cancer following estrogen ablation. Cancer Res 51:162–166Google Scholar
  34. 34.
    Redondo M, Villar E, Torres-Munoz J, Tellez T, Morell M, Petito CK (2000) Overexpression of clusterin in human breast carcinoma. Am J Pathol 157:393–399Google Scholar
  35. 35.
    Wellmann A, Thieblemont C, Pittaluga S, Sakai A, Jaffe ES, Siebert P, Raffeld M (2000) Detection of differentially expressed genes in lymphomas using cDNA arrays: identification of clusterin as a new diagnostic marker for anaplastic large-cell lymphomas. Blood 96:398–404Google Scholar
  36. 36.
    Parczyk K, Pilarsky C, Rachel U, Koch-Brandt C (1994) Gp80 (clusterin; TRPM-2) mRNA level is enhanced in human renal clear cell carcinomas. J Cancer Res Clin Oncol 120:186–188Google Scholar
  37. 37.
    Montpetit ML, Lawless KR, Tenniswood M (1986) Androgen-repressed messages in the rat ventral prostate. Prostate 8:25–36Google Scholar
  38. 38.
    Budendorf L, Kolmer M, Kononen J et al. (1999) Hormone therapy failure in human prostate cancer: analysis by complementary DNA and tissue arrays. J Natl Cancer Inst 91:1758–1764CrossRefGoogle Scholar
  39. 39.
    Kyprianou N, English HF, Isaacs JT (1990) Programmed cell death during regression of PC-82 human prostate cancer following androgen ablation. Cancer Res 50:3748–3753Google Scholar
  40. 40.
    Tenniswood MP, Guenette RS, Lakins J, Mooibroek M, Wong P, Welsh JE (1992) Active cell death in hormone-dependent tissues. Cancer Metastasis Rev 11:197–220CrossRefGoogle Scholar
  41. 41.
    Danik M, Chabot JG, Mercier C, Benabid AL, Chauvin C, Quirion R, Suh M (1991) Human gliomas and epileptic foci express high levels of a mRNA related to rat testicular sulfated glycoprotein 2, a purported marker of cell death. Proc Natl Acad Sci U S A 88:8577–8581Google Scholar
  42. 42.
    Connor J, Buttyan R, Olsson CA, D’Agati V, O’Toole K, Sawczuk IS (1991) SGP-2 expression as a genetic marker of progressive cellular pathology in experimental hydronephrosis. Kidney Int 39:1098–1103Google Scholar
  43. 43.
    Ho SM, Leav I, Ghatak S, Merk F, Jagannathan VS, Mallery K (1998) Lack of association between enhanced TRPM-2/clusterin expression and increased apoptotic activity in sex-hormone-induced prostatic dysplasia of the Noble rat. Am J Pathol 153:131–139Google Scholar
  44. 44.
    Schwochau GB, Nath KA, Rosenberg ME (1998) Clusterin protects against oxidative stress in vitro through aggregative and nonaggregative properties. Kidney Int 53:1647–1653CrossRefPubMedGoogle Scholar
  45. 45.
    French LE, Sappino AP, Tschopp J, Schifferli JA (1992) Distinct sites of production and deposition of the putative cell death marker clusterin in the human thymus. J Clin Invest 90:1919–1925Google Scholar
  46. 46.
    Yang CR, Leskov K, Hosley-Eberlein K, Criswell T, Pink JJ, Kinsella TJ, Boothman DA (2000) Nuclear clusterin/XIP8, an x-ray-induced Ku70-binding protein that signals cell death. Proc Natl Acad Sci U S A 97:5907–5912Google Scholar
  47. 47.
    Koch-Brandt C, Morgans C (1996) Clusterin: a role in cell survival in the face of apoptosis? Prog Mol Subcell Biol 16:130–149Google Scholar
  48. 48.
    Michel D, Chatelain G, North S, Brun G (1997) Stress-induced transcription of the clusterin/apoJ gene. Biochem J 328:45–50PubMedGoogle Scholar
  49. 49.
    Miyake H, Rennie P, Nelson C, Gleave ME (2000) Acquisition of chemoresistant phenotype by overexpression of the antiapoptotic gene, testosterone-repressed prostate message-2 (TRPM-2), in prostate cancer xenograft models. Cancer Res 60:2547–2554Google Scholar
  50. 50.
    Zellweger T, Chi K, Miyake H, Adomat H, KiyamaS, Skov K, Gleave M (2002) Enhanced radiation sensitivity in prostate cancer by inhibition of the cell survival protein clusterin. Clin Cancer Res 8:3276–3284Google Scholar
  51. 51.
    Criswell T, Klokov D, Beman M, Lavik JP, Boothman DA (2003) Repression of IR-inducible clusterin expression by the p53 tumour suppression protein. Cancer Biol Ther 2:372–380Google Scholar
  52. 52.
    Crescioli C, Ferruzzi P, Caporali A, Scaltriti M, Bettuzzi S, Mancina R, Gelmini S, Serio M, Villari D, Vannelli GB, Colli E, Adorini L, Maggi M (2004) Inhibition of prostate cell growth by BXL-628, a calcitriol analogue selected for a phase II clinical trial in patients with benign prostate hyperplasia. Eur J Endocrinol 150: 591–603Google Scholar
  53. 53.
    Jones SE, Jomary C (2002) Clusterin. IntJBiochem Cell Biol 34:427–431Google Scholar
  54. 54.
    Wilson MR, Easterbrook-Smith SB (2000) Clusterin is a secreted mammalian chaperone. Trends Biochem Sci 25:95–98CrossRefPubMedGoogle Scholar
  55. 55.
    Humphreys DT, Carver JA, Easterbrook-Smith SB, Wilson MR (1999) Clusterin has chaperone-like activity similar to that of small heat shock proteins. J Biol Chem 274:6875–6881CrossRefGoogle Scholar
  56. 56.
    Boise LH, Gonzalez-Garcia M, Postema CE, Ding L, Lindsten T, Turka LA, Mao X, Nunez G, Thompson CB (1993) bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell 74:597–608Google Scholar
  57. 57.
    Rouayrenc JF, Boise LH, Thompson CB, Privat A, Patey G (1995) Presence of the long and the short forms of Bcl-X in several human and murine tissues. C R Acad Sci III 318:537–540Google Scholar
  58. 58.
    Minn AJ, Boise LH, Thompson CB (1996) Bcl-x(S) anatagonizes the protective effects of Bcl-x(L). J Biol Chem 271:6306–6312Google Scholar
  59. 59.
    Sintich SM, Steinberg J, Kozlowski JM, Lee C, Pruden S, Sayeed S, Sensibar JA (1999) Cytotoxic sensitivity to tumor necrosis factor alpha in PC3 and LNCaP prostatic cancer cells is regulated by extracellular levels of SGP-2 (clusterin). Prostate 39:87–93Google Scholar
  60. 60.
    Miyake H, Hara I, Kamidono S, Gleave ME, Eto H (2003) Resistance to cytotoxic chemotherapy-induced apoptosis in human prostate cancer cells is associated with intracellular clusterin expression. Oncol Rep 10:469–473Google Scholar
  61. 61.
    Wong P, Pineault J, Lakins J, Taillefer D, Leger J, Wang C, Tenniswood M (1993) Genomic organization and expression of the rat TRPM-2 (clusterin) gene, a gene implicated in apoptosis. J Biol Chem 268:5021–5031Google Scholar
  62. 62.
    Lakins J, Bennett SA, Chen JH, Arnold JM, Morrissey C, Wong P, O’Sullivan J, Tenniswood M (1998) Clusterin biogenesis is altered during apoptosis in the regressing rat ventral prostate. J Biol Chem 273:27887–27895Google Scholar
  63. 63.
    Kounnas MZ, Loukinova EB, Stefansson S, Harmony JA, Brewer BH, Strickland DK, Argraves WS (1995) Identification of glycoprotein 330 as an endocytic receptor for apolipoprotein J/clusterin. J Biol Chem 270:13070–13075Google Scholar
  64. 64.
    Sensibar JA, Sutkowski DM, Raffo A et al. (1995) Prevention of cell death induced by tumor necrosis factor in alpha LNCaP cells by overexpression of sulfated glycoprotein-2 (clusterin). Cancer Res 55:2431–2437Google Scholar
  65. 65.
    Miyake H, Hara I, Gleave ME, Eto H (2004) Protection of androgen-dependent human prostate cancer cells from oxidative stress-induced DNA damage by overexpression of clusterin and its modulation by androgen. Prostate 61: 318–323Google Scholar
  66. 66.
    Pins MR, Fiadjoe JE, Korley F, Wong M, Rademaker AW, Jovanovic B, Yoo TK, Kozlowski JM, Raji A, Yang XJ, Lee C (2004) Clusterin as a possible predictor for biochemical recurrence of prostate cancer following radical prostatectomy with intermediate Gleason scores: a preliminary report. Prostate Cancer Prostatic Dis 7:243–248Google Scholar
  67. 67.
    July L, Akbari M, Zellweger T, Jones EC, Goldenberg SL, Gleave ME (2002) Clusterin expression is significantly enhanced in prostate cancer cells following androgen withdrawal therapy. Prostate 50:179–188CrossRefGoogle Scholar
  68. 68.
    Miyake H, Hara I, Kamidono S, Gleave ME (2001) Synergistic chemosensitization and inhibition of tumour growth and metastasis by the antisense oligodeoxynucleotide targeting clusterin gene in a human bladder cancer model. Clin Cancer Res 7:4245–4252Google Scholar
  69. 69.
    Zellweger T, Miyake H, July L, Akbari M, Kiyama S, Gleave ME (2001) Chemosensitization of a human renal cell cancer model by antisense TRPM-2 oligodeoxynucleotides both in vitro and in vivo. Neoplasia 3:1–8Google Scholar
  70. 70.
    July LV, Beraldi E, So AI, Evans K, English J, Fazli L, Gleave ME (2004) Nucleotide-based therapies targeting clusterin chemosensitizes human lung adenocarcinoma cells both in vitro and in vivo. Mol Cancer Ther 3:223–232Google Scholar
  71. 71.
    Crooke ST (1993) Therapeutic applications of oligonucleotides. Annu Rev Pharmacol Toxicol 32:329–376Google Scholar
  72. 72.
    Saijo Y, Perlaky L, Wang H, Busch H (1994) Pharmacokinetics, tissue distribution, and stability of antisense oligodeoxynucleotide phosphorothioate ISIS 3466 in mice. Oncol Res 6:243–249Google Scholar
  73. 73.
    Monia BP, Johnston JF, Geiger T, Muller M, Fabbro D (1996) Antitumor activity of a phosphorothioate antisense oligodeoxynucleotide targeted against C-raf kinase. Nat Med 2:668–675Google Scholar
  74. 74.
    Zellweger T, Miyake H, Cooper S, Chi K, Conklin BS, Monia B, Gleave ME (2001) Antitumor activity of antisense clusterin oligonucleotides is improved in vitro and in vivo by incorporation of 2′-o-(2-methoxy) ethyl chemistry. J Pharmacol Exp Ther 298:934–940Google Scholar
  75. 75.
    Henry S, Stecker K, Brooks D, Monteith D, Conklin B, Bennett CF (2000) Chemically modified oligonucleotides exhibit decreased immune stimulation in mice. J Pharmacol Exp Ther 292:468–479Google Scholar
  76. 76.
    Miyake H, Chi K, Gleave ME (2000) Antisense TRPM-2 oligodeoxynucleotides chemosensitize human androgen-independent PC-3 prostate cancer cells both in vitro and in vivo. Clin Cancer Res 6:1655–1663Google Scholar
  77. 77.
    Trougakso IP, So A, Jansen B, Gleave ME, Gonos ES (2004) Silencing expression of the clusterin/apolipoprotein J gene in human cancer cells using small interfering RNA induces spontaneous apoptosis reduced growth ability, and cell sensitization to genotoxic and oxidative stress. Cancer Res 64:1834–1842Google Scholar
  78. 78.
    Kyprianou N, King ED, Bradbury D, Rhee JG (1997) Bcl-2 overexpression delays radiation-induced apoptosis without affecting the clonogenic survival of human prostate cancer cells. Int J Cancer 70:341–348Google Scholar
  79. 79.
    Zellweger T, Kiyama S, Chi K, Miyake H, Adomat H, Skov K, Gleave ME (2003) Overexpression of the cytoprotective protein clusterin decreases radiosensitivity in the human LNCaP prostate tumour model. BJU Int 92:463–469Google Scholar
  80. 80.
    Chi KN, Eisenhauer E, Fazli L, Jones EC, Powers J, Ayers D, Goldenberg SL, Gleave ME (2004) A phase I pharmacokinetic (PK) and pharmacodynamic (PD) study of OGX-011, a 2′methoxyethyl phosphorothioate antisense to clusterin, in patients with prostate cancer prior to radical prostatectomy. J Clin Ocol, Proceedings of American Society of Clinical Oncology, 2004Google Scholar
  81. 81.
    Hoeller C, Pratscher B, Thallinger C, Winter D, Fink D, Kovacic B, Sexl V, Wacheck V, Gleave ME, Pehamberger H, Jansen B (2005) Clusterin regulates drug-resistance in melanoma cells.J Invest Dermatol (in press)Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Division of UrologyUniversity of British ColumbiaVancouverCanada
  2. 2.The Prostate CentreVancouver General HospitalVancouverCanada

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