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Medical Oncology

, Volume 27, Issue 3, pp 592–599 | Cite as

Treatment of prostate and breast tumors employing mono- and bi-specific antisense oligonucleotides targeting apoptosis inhibitory proteins clusterin and bcl-2

  • Marvin RubensteinEmail author
  • Paulus Tsui
  • Patrick Guinan
Original Paper

Abstract

Antisense oligonucleotides (oligos) have demonstrated their efficacy in inhibiting the growth of prostate and breast tumor cells. Previous studies employed first generation, phosphorothioated, cDNA oligos synthesized complimentary to mRNA encoding transforming growth factor-alpha (TGF-α), epidermal growth factor receptor (EGFR), the anti-apoptosis protein bcl-2, and the androgen receptor (AR). In an effort to construct oligos with greater than one mRNA binding site, bi-specifics have been developed which target combinations of the above proteins, and these have been shown at least as effective as the mono-specific oligos from which their sequences were derived. While all bi-specifics have inhibitory effects, which can be enhanced by the combined administration of an additional chemotherapeutic agent, those bi-specifics which target bcl-2 and EGFR were reported to be the most effective. The experiments presented here are an effort to evaluate a new group of bi-specifics whose targets include the chaperone protein clusterin, whose expression is up regulated in many tumors and activity is known to inhibit apoptosis. Of particular interest were those bi-specifics constructed to target both clusterin and bcl-2 (also an apoptosis inhibitory protein). Cell lines targeted included both prostate LNCaP and PC-3, as well as the breast derived MCF-7. In order to identify agents which enhance oligo activity, but contribute less toxicity, oligos were tested both alone and in combination with either the immune inhibitor Rapamycin, or the chemotherapeutic (and more toxic) Taxol. Results indicate that bi-specifics targeting clusterin are statistically effective, and are similarly enhanced by Rapamycin, or Taxol. When bi-specifics including clusterin as a target, were tested against LNCaP and MCF-7 cells, the level of activity was intermediate between that of the mono-specific compounds tested separately. In experiments which compared both, bi-specifics which included a target for clusterin had inhibitory activity similar to the previously described bi-specifics directed towards bcl-2 and EGFR.

Keywords

Antisense Prostate cancer Breast cancer Therapy 

Notes

Acknowledgments

The Cellular Biology laboratory at the Hektoen Institute is supported, in part, by the Blum Kovler Foundation, the Cancer Federation, Safeway/Dominicks Campaign for Breast Cancer Awareness, Lawn Manor Beth Jacob Hebrew Congregation, the Max Goldenberg Foundation, the Sternfeld Family Foundation, and the Herbert C. Wenske Foundation.

References

  1. 1.
    Rubenstein M, Mirochnik Y, Chow P, Guinan P. Antisense oligonucleotide intralesional therapy of human PC-3 prostate tumors carried in athymic nude mice. J Surg Oncol. 1996;62:194–200. doi: 10.1002/(SICI)1096-9098(199607)62:3<194::AID-JSO9>3.0.CO;2-2.CrossRefPubMedGoogle Scholar
  2. 2.
    Rubenstein M, Muchnik S, Dunea G, Chous P, Guinan P. Inoculation of prostatic tumors with antisense oligonucleotides against mRNA encoding growth factors and receptors. In: Einhorn J, Nord CK, Norby SR, editors. Recent advances in chemotherapy. Washington, DC: American Society of Microbiology; 1994. p. 898–9.Google Scholar
  3. 3.
    Rubenstein M, Mirochnik Y, Chou P, Guinan P. Growth factor deprivation therapy of hormone insensitive prostate and breast cancers utilizing antisense oligonucleotides. Meth Find Clin Pharmacol. 1998;20:825–31. doi: 10.1358/mf.1998.20.10.487534.CrossRefGoogle Scholar
  4. 4.
    Rubenstein M, Tsui P, Guinan P. Treatment of MCF-7 breast cancer cells employing mono- and bispecific antisense oligonucleotides having binding specificity toward proteins associated with autocrine regulated growth and bcl-2. Med Oncol. 2008;25:182–6. doi: 10.1007/s12032-007-9018-y.CrossRefPubMedGoogle Scholar
  5. 5.
    Rubenstein M, Glick R, Lichtor T, Mirochnik Y, Chou P, Guinan P. Treatment of the T98G glioblastoma cell line with antisense oligonucleotides directed toward mRNA encoding transforming growth factor-alpha and the epidermal growth factor receptor. Med Oncol. 2001;18:121–30. doi: 10.1385/MO:18:2:121.CrossRefPubMedGoogle Scholar
  6. 6.
    Rubenstein M, Tsui P, Guinan P. Construction of a bispecific antisense oligonucleotide containing multiple binding sites for the treatment of hormone insensitive prostate tumors. Med Hypotheses. 2005;65:905–7. doi: 10.1016/j.mehy.2004.12.032.CrossRefPubMedGoogle Scholar
  7. 7.
    Rubenstein M, Tsui P, Guinan P. Bispecific antisense oligonucleotides with multiple binding sites for the treatment of prostate tumors and their applicability to combination therapy. Methods Find Clin Pharmacol. 2006;28:1–4.Google Scholar
  8. 8.
    Rubenstein M, Tsui P, Guinan P. Bispecific antisense oligonucleotides having binding sites directed against an autocrine regulated growth pathway and bcl-2 for the treatment of prostate tumors. Med Oncol. 2007;24:189–96. doi: 10.1007/BF02698039.CrossRefPubMedGoogle Scholar
  9. 9.
    Zhang H, Kim JK, Edwards CA, Xy Z, Taichman R, Wang C-Y. Clusterin inhibits apoptosis by interacting with activated Bax. Nature Cell Biol. 2005;7:909–15. doi: 10.1038/ncb1291.CrossRefPubMedGoogle Scholar
  10. 10.
    Redondo M, Téllez T, Roldan MJ, Serrano A, García-Aranda M, Gleave ME, et al. Anticlusterin treatment of breast cancer cells increases the sensitivities of chemotherapy and tamoxifen and counteracts the inhibitory action of dexamethasone on chemotherapy-induced cytotoxicity. Breast Cancer Res. 2007;9:R86. doi: 10.1186/bcr1835.CrossRefPubMedGoogle Scholar
  11. 11.
    Rubenstein M, Tsui P, Guinan P. Multigene targeting of signal transduction pathways for the treatment of breast and prostate tumors. Comparisons between combination therapies employing bispecific oligonucleotides with either Rapamycin or Paclitaxel. Med Oncol. 2009;26:124.Google Scholar
  12. 12.
    Rubenstein M, Anderson KM, Tsui P, Guinan P. Synthesis of branched antisense oligonucleotides having multiple specificities: treatment of hormone insensitive prostate cancer. Med Hypotheses. 2006;67:1374–9. doi: 10.1016/j.mehy.2006.05.055.CrossRefGoogle Scholar
  13. 13.
    Weinberg R. The biology of cancer. 16.15 mTOR, a master regulator of cell physiology, represents an attractive target for anti-cancer therapy. New York: Garland Science; 2007. p. 782–787.Google Scholar
  14. 14.
    Sabatini DM. mTOR and cancer: insights into a complex relationship. Nat Rev Cancer. 2006;6:729–34. doi: 10.1038/nrc1974.CrossRefPubMedGoogle Scholar
  15. 15.
    Shaw RJ, Cantley LC. Ras, PI(3)K and mTOR signaling controls tumour cell growth. Nature. 2006;441:424–30. doi: 10.1038/nature04869.CrossRefPubMedGoogle Scholar
  16. 16.
    Amornphimoltham P, Patel V, Leelahavanichkul K, Abraham RT, Gutkind JS. A retroinhibition approach reveals a tumor cell-autonomous response to Rapamycin in head and neck cancers. Cancer Res. 2008;68:1144–53. doi: 10.1158/0008-5472.CAN-07-1756.CrossRefPubMedGoogle Scholar
  17. 17.
    Sanfilippo NJ, Taneja SS, Chachoua A, Lepor H, Formenti SC. Phase I/II study of biweekly paclitaxel and radiation in androgen-ablated locally advanced prostate cancer. J Clin Oncol. 2008;26:2973–8. doi: 10.1200/JCO.2007.14.4105.CrossRefPubMedGoogle Scholar
  18. 18.
    Gleave M, Tolcher A, Miyake H, Nelson C, Brown B, Beraldi E, et al. Progression to androgen independence is delayed by adjuvant treatment with antisense bcl-2 oligodeoxynucleotides after castration in the LNCaP prostate tumor model. Clin Cancer Res. 1999;5:2891–8.PubMedGoogle Scholar
  19. 19.
    Rubenstein M, Slobodskoy L, Mirochnik Y, Guinan P. Inhibition of PC-3 prostate cancer cell growth in vitro using both antisense oligonucleotides and Taxol. Med Oncol. 2003;20:29–35. doi: 10.1385/MO:20:1:29.CrossRefPubMedGoogle Scholar
  20. 20.
    Tsui P, Rubenstein M, Guinan P. Synergistic effects of combination therapy employing antisense oligonucleotides with traditional chemotherapeutics in the PC-3 prostate cancer model. Med Oncol. 2004;21:339–48. doi: 10.1385/MO:21:4:339.CrossRefPubMedGoogle Scholar
  21. 21.
    Fluiter K, Frieden M, Vreijling J, Jakobs M., Rosenbohm C, Koch T, Baas F. The properties of novel generation LNA chemistries in antisense oligonucleotides. Effects on biodistribution and efficacy of tumor growth inhibition in vivo. Proc Am Assoc Cancer Res. 2004; 45:Abst 2931.Google Scholar
  22. 22.
    Janus A, Robak T, Smolewski P. The mammalian target of the Rapamycin (mTOR) kinase pathway; its role in tumourigenesis and target antitumour therapy. Cell Mol Biol Lett. 2005;10:479–97.PubMedGoogle Scholar
  23. 23.
    Hudson CC, Liu M, Chiang GG, Otterness DM, Loomis DC, Kaper F, et al. Regulation of hypoxia-inducible factor 1α expression and function by the mammalian target of Rapamycin. Mol Cell Biol. 2002;22:7004–14. doi: 10.1128/MCB.22.20.7004-7014.2002.CrossRefPubMedGoogle Scholar
  24. 24.
    Atkins MB, Hidalgo MK, Stadler WM, Logan TF, Dutcher JP, Hudes GR, et al. Randomized phase II study of multiple dose levels of CCI-779, a novel mammalian target of Rapamycin kinase inhibitor, in patients with advanced refractory renal cell carcinoma. J Clin Oncol. 2004;22:909–18. doi: 10.1200/JCO.2004.08.185.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2009

Authors and Affiliations

  • Marvin Rubenstein
    • 1
    • 2
    • 3
    • 4
    Email author
  • Paulus Tsui
    • 1
    • 2
  • Patrick Guinan
    • 1
    • 2
    • 4
    • 5
  1. 1.Division of Cellular BiologyHektoen Institute for Medical ResearchChicagoUSA
  2. 2.Division of UrologyStroger Hospital of Cook CountyChicagoUSA
  3. 3.Departments of BiochemistryRush University Medical CenterChicagoUSA
  4. 4.Department of UrologyRush University Medical CenterChicagoUSA
  5. 5.Department of UrologyUniversity of Illinois at ChicagoChicagoUSA

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