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The topoisomerase I inhibitor topotecan increases the sensitivity of prostate tumor cells to TRAIL/Apo-2L-induced apoptosis

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Abstract

Purpose

TRAIL/Apo-2L is cytotoxic against numerous prostate tumor cell lines; however, some lines are more resistant than others. Identification of an agent that increases prostate tumor cell sensitivity to TRAIL/Apo-2L would prove valuable for TRAIL/Apo-2L-mediated tumor therapy. Thus, we examined the effect of combining five clinically approved chemotherapeutic agents with TRAIL/Apo-2L for treating prostate tumor cells.

Methods

Four human prostate tumor cell lines were initially tested for TRAIL/Apo-2L sensitivity. Subsequent studies examined whether the TRAIL/Apo-2L-induced killing of DU-145 cells was augmented in the presence of the chemotherapeutic molecules, as measured by annexin V-FITC/propidium iodide staining. Furthermore, caspase 8 activation and BID cleavage were examined by immunoblotting. RT-PCR and flow cytometry were performed to monitor TRAIL-R1 and TRAIL-R2 levels after chemotherapeutic treatment.

Results

DU-145 cells were the least responsive of the prostate tumor cell lines tested to TRAIL/Apo-2L-induced death. Surprisingly, only topotecan, a topoisomerase I inhibitor, when used in combination with rTRAIL/Apo-2L led to significant apoptosis of DU-145 cells, as measured by caspase 8 activation, BID cleavage, and annexin V-FITC/PI staining. Topotecan alone had little to no toxicity on the DU-145 cells. Furthermore, the increase in TRAIL/Apo-2L sensitivity following topotecan treatment correlated with increased expression of TRAIL-R1 and TRAIL-R2 and decreased intracellular levels of the antiapoptotic protein survivin.

Conclusions

Our results define a promising direction for alternative therapies against androgen-independent prostate cancers. The sensitivity of DU-145 cells to TRAIL/Apo-2L was dramatically increased when combined with topotecan, suggesting that low-dose topotecan treatment to upregulate TRAIL-R1 and TRAIL-R2 and downregulate survivin, followed by TRAIL/Apo-2L administration, may be a viable therapy for treating cancer of the prostate.

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Abbreviations

BID:

Bcl-2-interacting domain death agonist

FADD:

Fas-associated death domain

FLIP:

FADD-like IL-1β-converting enzyme-inhibitory protein

IAP:

Inhibitor of apoptosis protein

MDR:

Multidrug resistance

PrEC:

Prostate epithelial cells

RT-PCR:

Reverse transcription-polymerase chain reaction

TRAIL:

TNF-related apoptosis-inducing ligand

References

  1. Ambrosini G, Adid A, Altieri D (1997) A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 3:917

    CAS  PubMed  Google Scholar 

  2. Armitage RJ (1994) Tumor necrosis factor receptor superfamily members and their ligands. Curr Opin Immunol 6:407

    CAS  PubMed  Google Scholar 

  3. Ashkenazi A, Pai RC, Fong S, Leung S, Lawrence DA, Marsters SA, Blackie C, Chang L, McMurtrey AE, Hebert A, DeForge L, Koumenis IL, Lewis D, Harris L, Bussiere J, Koeppen H, Shahrokh Z, Schwall RH (1999) Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 104:155

    CAS  PubMed  Google Scholar 

  4. Berenbaum MC (1981) Criteria for analyzing interactions between biologically active agents. Adv Cancer Res 35:269

    CAS  PubMed  Google Scholar 

  5. Burden DA, Kingma PS, Froelich-Ammon SJ, Bjornsti MA, Patchan MW, Thompson RB, Osheroff N (1996) Topoisomerase II.etoposide interactions direct the formation of drug-induced enzyme-DNA cleavage complexes. J Biol Chem 271:29238

    Article  CAS  PubMed  Google Scholar 

  6. Chen H, Liu X, Patel DJ (1996) DNA bending and unwinding associated with actinomycin D antibiotics bound to partially overlapping sites on DNA. J Mol Biol 258:457

    Article  CAS  PubMed  Google Scholar 

  7. Chinnaiyan AM, Prasad U, Shankar S, Hamstra DA, Shanaiah M, Chenevert TL, Ross BD, Rehemtulla A (2000) Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy. Proc Natl Acad Sci U S A 97:1754

    Article  CAS  PubMed  Google Scholar 

  8. Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27

    CAS  PubMed  Google Scholar 

  9. Cohen SM, Lippard SJ (2001) Cisplatin: from DNA damage to cancer chemotherapy. Prog Nucleic Acid Res Mol Biol 67:93

    CAS  PubMed  Google Scholar 

  10. Degli-Esposti MA, Dougall WC, Smolak PJ, Waugh JY, Smith CA, Goodwin RG (1997) The novel receptor TRAIL-R4 induces NFκB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain. Immunity 7:813

    CAS  PubMed  Google Scholar 

  11. Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, Henson PM (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol 148:2207

    CAS  PubMed  Google Scholar 

  12. Fulda S, Debatin KM (2002) IFN gamma sensitizes for apoptosis by upregulating caspase-8 expression through the Stat1 pathway. Oncogene 21:2295

    Article  CAS  PubMed  Google Scholar 

  13. Fulda S, Meyer E, Debatin KM (2002) Inhibition of TRAIL-induced apoptosis by Bcl-2 overexpression. Oncogene 21:2283

    Article  CAS  PubMed  Google Scholar 

  14. George JW, Ghate S, Matson SW, Besterman JM (1992) Inhibition of DNA helicase II unwinding and ATPase activities by DNA-interacting ligands. Kinetics and specificity. J Biol Chem 267:10683

    CAS  PubMed  Google Scholar 

  15. Griffith TS, Broghammer EL (2001) Suppression of tumor growth following intralesional therapy with TRAIL recombinant adenovirus. Mol Ther 4:257

    Article  CAS  PubMed  Google Scholar 

  16. Griffith TS, Lynch DH (1998) TRAIL: a molecule with multiple receptors and control mechanisms. Curr Opin Immunol 10:559

    CAS  PubMed  Google Scholar 

  17. Griffith TS, Chin WA, Jackson GC, Lynch DH, Kubin MZ (1998) Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells. J Immunol 161:2833

    CAS  PubMed  Google Scholar 

  18. Gross A, Yin XM, Wang K, Wei MC, Jockel J, Milliman C, Erdjument-Bromage H, Tempst P, Korsmeyer SJ (1999) Caspase cleaved BID targets mitochondria and is required for cytochrome c release, while BCL-XL prevents this release but not tumor necrosis factor-R1/Fas death. J Biol Chem 274:1156

    CAS  PubMed  Google Scholar 

  19. Grossman D, McNiff JM, Li F, Altieri DC (1999) Expression and targeting of the apoptosis inhibitor, survivin, in human melanoma. J Invest Dermatol 113:1076

    Article  CAS  PubMed  Google Scholar 

  20. Guan B, Yue P, Clayman GL, Sun S-Y (2001) Evidence that the death receptor DR4 is a DNA damage-inducible, p53-regulated gene. J Cell Physiol 188:98

    Article  CAS  PubMed  Google Scholar 

  21. Guan B, Yue P, Lotan R, Sun S-Y (2002) Evidence that the human death receptor 4 is regulated by activator protein 1. Oncogene 21:3121

    Article  CAS  PubMed  Google Scholar 

  22. Hawkins CJ, Vaux DL (1997) The role of the Bcl-2 family of apoptosis regulatory proteins in the immune system. Semin Immunol 9:2533

    Article  Google Scholar 

  23. Hendricks CB, Rowinsky EK, Grochow LB, Donehower RC, Kaufmann SH (1992) Effect of P-glycoprotein expression on the accumulation and cytotoxicity of topotecan (SK&F 104864), a new camptothecin analogue. Cancer Res 52:2268

    CAS  PubMed  Google Scholar 

  24. Huennekens FM (1994) The methotrexate story: a paradigm for development of cancer chemotherapeutic agents. Adv Enzyme Regul 34:397

    Article  CAS  PubMed  Google Scholar 

  25. Jemal A, Thomas A, Murray T, Thun M (2002) Cancer statistics, 2002. CA Cancer J Clin 52:23

    PubMed  Google Scholar 

  26. Kischkel FC, Lawrence DA, Chuntharapai A, Schow P, Kim KJ, Ashkenazi A (2000) Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5. Cell 12:611

    CAS  Google Scholar 

  27. Kollmannsberger C, Mross K, Jakob A, Kanz L, Bokemeyer C (1999) Topotecan—a novel topoisomerase I inhibitor: pharmacology and clinical experience. Oncology 56:1

    CAS  PubMed  Google Scholar 

  28. Martin SJ, Reutelingsperger CPM, McGahon AJ, Rader JA, van Schie RCAA, LaFace DM, Green DR (1995) Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 182:1545

    CAS  PubMed  Google Scholar 

  29. Muzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J, Scaffidi C, Bretz JD, Zhang M, Gentz R, Mann M, Krammer PH, Peter ME, Dixit VM (1996) FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 85:817

    CAS  PubMed  Google Scholar 

  30. Nesterov A, Lu X, Johnson M, Miller GJ, Ivashchenko Y, Kraft AS (2001) Elevated AKT activity protects the prostate cancer cell line LNCaP from TRAIL-induced apoptosis. J Biol Chem 276:10767

    Article  CAS  PubMed  Google Scholar 

  31. Nimmanapalli R, Perkins CL, Orlando M, O'Bryan E, Nguyen D, Bhalla KN (2001) Pretreatment with paclitaxel enhances apo-2 ligand/tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis of prostate cancer cells by inducing death receptors 4 and 5 protein levels. Cancer Res 61:759

    CAS  PubMed  Google Scholar 

  32. Pan G, Ni J, Wei Y-F, Yu G-I, Gentz R, Dixit VM (1997) An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 277:815

    CAS  PubMed  Google Scholar 

  33. Pan G, O'Rourke K, Chinnaiyan AM, Gentz R, Ebner R, Ni J, Dixit VM (1997) The receptor for the cytotoxic ligand TRAIL. Science 276:111

    CAS  PubMed  Google Scholar 

  34. Pitti RM, Marsters SA, Ruppert S, Donahue CJ, Moore A, Ashkenazi A (1996) Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family. J Biol Chem 271:12687

    CAS  PubMed  Google Scholar 

  35. Shin MS, Kim HS, Lee SH, Park WS, Kim SY, Park JY, Lee JH, Lee SK, Lee SN, Jung SS, Han JY, Kim H, Lee JY, Yoo NJ (2001) Mutations of tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAIL-R1) and receptor 2 (TRAIL-R2) genes in metastatic breast cancers. Cancer Res 61:4942

    CAS  PubMed  Google Scholar 

  36. Sprick MR, Weigand MA, Rieser E, Rauch CT, Juo P, Blenis J, Krammer PH, Walczak H (2000) FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity 12:599

    CAS  PubMed  Google Scholar 

  37. Swana HS, Grossman D, Anthony JN, Weiss RM, Altieri DC (1999) Tumor content of the anti-apoptosis molecule survivin and recurrence of bladder cancer. N Engl J Med 341:452

    CAS  PubMed  Google Scholar 

  38. Tamm I, Wang Y, Sausville E, Scudiero DA, Vigna N, Oltersdorf T, Reed JC (1999) IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs. Cancer Res 58:5315

    Google Scholar 

  39. Tschopp J, Irmler M, Thome M (1998) Inhibition of Fas death signals by FLIPs. Curr Opin Immunol 10:552

    Article  PubMed  Google Scholar 

  40. Van Brussel JP, Jan Van Steenbrugge G, Van Krimpen C, Bogdanowicz JF, Van Der Kwast TH, Schroder FH, Mickisch GH (2001) Expression of multidrug resistance related proteins and proliferative activity is increased in advanced clinical prostate cancer. J Urol 165:130

    PubMed  Google Scholar 

  41. Waterhouse NJ, Goldstein JC, von Ahsen O, Schuler M, Newmeyer DD, Green DR (2001) Cytochrome c maintains mitochondrial transmembrane potential and ATP generation after outer mitochondrial membrane permeabilization during the apoptotic process. J Cell Biol 153:319

    Article  CAS  PubMed  Google Scholar 

  42. Wiley SR, Schooley K, Smolak PJ, Din WS, Huang C-P, Nicholl JK, Sutherland GR, Smith TD, Rauch C, Smith CA, Goodwin RG (1995) Identification and characterization of a new member of the TNF family that induces apoptosis. Immunity 3:673

    CAS  PubMed  Google Scholar 

  43. Wu GS, Burns TF, McDonald ER 3rd, Jiang W, Meng R, Krantz ID, Kao G, Gan DD, Zhou JY, Muschel R, Hamilton SR, Spinner NB, Markowitz S, Wu G, el-Deiry WS (1997) KILLER/DR5 is a DNA damage-inducible p53-regulated death receptor gene. Nat Genet 17:141

    CAS  PubMed  Google Scholar 

  44. Wu XX, Kakehi Y, Mizutani Y, Kamoto T, Kinoshita H, Isogawa Y, Terachi T, Ogawa O (2002) Doxorubicin enhances TRAIL-induced apoptosis in prostate cancer. Int J Oncol 20:949

    CAS  PubMed  Google Scholar 

  45. Yoshida T, Maeda A, Tani N, Sakai T (2001) Promoter structure and transcription initiation sites of the human death receptor 5/TRAIL-R2 gene. FEBS Lett 507:381

    Article  CAS  PubMed  Google Scholar 

  46. Yu R, Mandlekar S, Ruben S, Ni J, Kong AN (2000) Tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in androgen-independent prostate cancer cells. Cancer Res 60:2384

    CAS  PubMed  Google Scholar 

  47. Zisman A, Ng CP, Pantuck AJ, Bonavida B, Belldegrun AS (2001) Actinomycin D and gemcitabine synergistically sensitize androgen-independent prostate cancer cells to Apo2L/TRAIL-mediated apoptosis. J Immunother 24:459

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Drs. Ben Elzey and Tim Ratliff for careful reading the manuscript. We would also like to acknowledge Linda Buckner for secretarial assistance. This work was supported by an American Cancer Society Institutional Seed Grant (no. IN-122U) administered through the University of Iowa's Holden Comprehensive Cancer Center, the Edwin Beer Program of The New York Academy of Medicine, and a Department of Defense Prostate Cancer Research Program New Investigator Award (PC010599).

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Correspondence to Thomas S. Griffith.

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Griffith, T.S., Kemp, T.J. The topoisomerase I inhibitor topotecan increases the sensitivity of prostate tumor cells to TRAIL/Apo-2L-induced apoptosis. Cancer Chemother Pharmacol 52, 175–184 (2003). https://doi.org/10.1007/s00280-003-0656-2

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