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CLL

Inhibition of RNA transcription: a biochemical mechanism of action against chronic lymphocytic leukemia cells by fludarabine

Leukemia volume 14, pages 1405–1413 (2000)Cite this article

Abstract

Fludarabine is a nucleotide analog effective in the clinical treatment of chronic lymphocytic leukemia (CLL) and other indolent lymphocytic malignancies. Although the incorporation of fludarabine into DNA is a key event in causing cytotoxicity in proliferating leukemia cells, the precise mechanisms by which fludarabine kills CLL cells remain unclear because of the quiescent nature of this malignancy. The present study demonstrated that inhibition of RNA transcription correlated significantly with the cytotoxic action of fludarabine in CLL cells. In contrast, suppression of the low level of DNA synthesis did not affect the survival of the leukemia cells. In addition, inhibition of fludarabine incorporation into cellular DNA through repair synthesis in CLL cells did not alter the cytotoxicity of this drug. Rather, inhibition of RNA synthesis by fludarabine led to a specific diminishment of certain cellular proteins from CLL cells. The combination of fludarabine with another RNA synthesis inhibitor, actinomycin D, or with the protein synthesis inhibitor, puromycin, substantially enhanced the cytotoxic activity against CLL cells. These results suggest that termination of mRNA transcription and the consequent depletion of proteins required for cell survival may be a novel biochemical mechanism of action of fludarabine in CLL cells. Thus, inhibition of RNA/protein synthesis may provide a new therapeutic strategy for the treatment of CLL patients.

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References

  1. Reed JC . Molecular biology of chronic lymphocytic leukemia Semin Oncol 1998 25: 11–18

    CAS  PubMed  Google Scholar 

  2. Kipps T J . Chronic lymphocytic leukemia Curr Opin Hematol 1997 4: 268–276

    PubMed  Google Scholar 

  3. Keating MJ, O'Brien S, Lerner S, Koller C, Beran M, Robertson LE, Freireich EJ, Estey E, Kantarjian H . Long-term follow-up of patients with chronic lymphocytic leukemia (CLL) receiving fludarabine regimens as initial therapy Blood 1998 92: 1165–1171

    CAS  PubMed  Google Scholar 

  4. Keating M J, O'Brien S, McLaughlin P, Dimopoulos M, Gandhi V, Plunkett W, Lerner S, Kantarjian H, Estey E . Clinical experience with fludarabine in hemato-oncology Hematol Cell Ther 1996 38: (Suppl. 2) 83–91

    Google Scholar 

  5. Adkins JC, Peters DH, Markham A . Fludarabine: an update of its pharmacology and use in the treatment of haematological malignancies Drugs 1997 53: 1005–1037

    CAS  PubMed  Google Scholar 

  6. Keating MJ, O'Brien S, Plunkett W, Robertson LE, Gandhi V, Estey E, Dimopoulos M, Cabanillas F, Kemena A, Kantarjian H . Fludarabine phosphate: a new active agent in hematologic malignancies Semin Hematol 1994 31: 28–39

    CAS  PubMed  Google Scholar 

  7. O'Brien S, Kantarjian H, Beran M, Koller C, Talpaz M, Lerner S, Keating MJ . Fludarabine and granulocyte colony-stimulating factor (G-CSF) in patients with chronic lymphocytic leukemia Leukemia 1997 11: 1631–1635

    CAS  PubMed  Google Scholar 

  8. Sorensen JM, Vena DA, Fallavollita A, Chun HG, Cheson BD . Treatment of refractory chronic lymphocytic leukemia with fludarabine phosphate via the group C protocol mechanism of the National Cancer Institute: five-year follow-up report J Clin Oncol 1997 15: 458–465

    CAS  PubMed  Google Scholar 

  9. Huang P, Chubb S, Plunkett W . Termination of DNA synthesis by 9-β-D-arabinofuranosyl-2-fluoroadenine: a mechanism for cytotoxicity J Biol Chem 1990 265: 16617–16625

    CAS  PubMed  Google Scholar 

  10. Huang P, Plunkett W . Fludarabine and gemcitabine-induced apoptosis: Incorporation of analogs into DNA is a critical event Cancer Chemother Pharmacol 1995 36: 181–188

    CAS  PubMed  Google Scholar 

  11. Huang P, Robertson LE, Wright S, Plunkett W . High molecular weight DNA fragmentation: a critical event in nucleoside analogue-induced apoptosis in leukemia cells Clin Cancer Res 1995 1: 1005–1013

    CAS  PubMed  Google Scholar 

  12. Huang P, Ballal K, Plunkett W . Biochemical characterization of the protein activity responsible for high molecular weight DNA fragmentation during drug-induced apoptosis Cancer Res 1997 57: 3407–3414

    CAS  PubMed  Google Scholar 

  13. Parker WB, Bapat AR, Shen JX, Townsend AJ, Cheng YC . Interaction of 2-halogenated dATP analogs (F, Cl, and Br) with human DNA polymerases, DNA primase, and ribonucleotide reductase Mol Pharmacol 1988 34: 485–491

    CAS  PubMed  Google Scholar 

  14. Catapano CV, Perrino FW, Fernandes DJ . Primer RNA chain termination induced by 9-β-D-arabinofuranosyl-2-fluoroadenine 5′-triphosphate: a mechanism of DNA synthesis inhibition J Biol Chem 1993 268: 7179–7185

    CAS  PubMed  Google Scholar 

  15. Yang SW, Huang P, Plunkett W, Becker FF, Chan JYH . Dual mode of inhibition of purified DNA ligase I from human cells by 9-β-D-arabinofuranosyl-2-fluoroadenine triphosphate J Biol Chem 1992 267: 2345–2349

    CAS  PubMed  Google Scholar 

  16. Huang P, Plunkett W . Action of 9-β-D-arabinofuranosyl-2-fluoroadenine on RNA metabolism Mol Pharmacol 1991 39: 449–455

    CAS  PubMed  Google Scholar 

  17. Foon KA, Rai KR, Gale RP . Chronic lymphocytic leukemia: new insights into biology and therapy Ann Intern Med 1990 113: 525–539

    CAS  PubMed  Google Scholar 

  18. Faguet GB . Chronic lymphocytic leukemia: an updated review J Clin Oncol 1994 12: 1974–1990

    CAS  PubMed  Google Scholar 

  19. Cheson BD, Bennett JM, Rai KR, Grever MR, Kay NE, Schiffer CA, Oken MM, Keating MJ, Boldt DH, Kempin SJ, Foon KA . Guidelines for clinical protocols for chronic lymphocytic leukemia: recommendations of the National Cancer Institute-sponsored working group Am J Hematol 1988 29: 152–163

    CAS  PubMed  Google Scholar 

  20. Xu Y-Z, Huang P, Plunkett W . Functional compartmentation of dCTP pools: preferential utilization of salvaged deoxycytidine for DNA repair in human lymphoblasts J Biol Chem 1995 270: 631–637

    CAS  PubMed  Google Scholar 

  21. Iwasaki H, Huang P, Keating MJ, Plunkett W . Differential incorporation of ara-C, gemcitabine, and fludarabine into replicating and repairing DNA in proliferating human leukemia cells Blood 1997 90: 270–278

    CAS  PubMed  Google Scholar 

  22. Robertson LE, Chubb S, Meyn RE, Story M, Ford R, Hittelman WN, Plunkett W . Induction of apoptotic cell death in chronic lymphocytic leukemia by 2-chloro-2′-deoxyadenosine and 9-β-D-arabinosyl-2-fluoroadenine Blood 1993 81: 143–150

    CAS  PubMed  Google Scholar 

  23. Cherrington JM, Allen SJ, McKee BH, Chen MS . Kinetic analysis of the interaction between the diphosphate of (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine, ddCTP, AZTTP, and FIAUTP with human DNA polymerases beta and gamma Biochem Pharmacol 1994 48: 1986–1988

    CAS  PubMed  Google Scholar 

  24. Bouayadi K, Hoffmann JS, Fons P, Tiraby M, Reynes JP, Cazaux C . Overexpression of DNA polymerase beta sensitizes mammalian cells to 2′,3′-deoxycytidine and 3′-azido-3′-deoxythymidine Cancer Res 1997 57: 110–116

    CAS  PubMed  Google Scholar 

  25. Huang P, Chubb S, Hertel LW, Grindey GB, Plunkett W . Action of 2′,2′-difluorodeoxycytidine on DNA synthesis Cancer Res 1991 51: 6110–6117

    CAS  PubMed  Google Scholar 

  26. Aivasashvilli VA, Beabealashvilli RS . Sequence-specific inhibition of RNA elongation by actinomycin D FEBS Lett 1983 160: 124–128

    CAS  PubMed  Google Scholar 

  27. Li L, Liu X, Glassman AB, Keating MJ, Plunkett W, Yang LY . Fludarabine triphosphate inhibits nucleotide excision repair of cisplatin-induced DNA adducts in vitro Cancer Res 1997 57: 1487–1494

    CAS  PubMed  Google Scholar 

  28. Sandoval A, Consoli U, Plunkett W . Fludarabine-mediated inhibition of nucleotide excision repair induces apoptosis in quiescent human lymphocytes Clin Cancer Res 1996 2: 1731–1741

    CAS  PubMed  Google Scholar 

  29. Yang LY, Li L, Keating MJ, Plunkett W . Arabinosylfluoroadenine augments cisplatin cytotoxicity and inhibits cisplatin-DNA cross-link repair Mol Pharmacol 1995 47: 1072–1079

    CAS  PubMed  Google Scholar 

  30. Spriggs D, Robbins G, Mitchell T, Kufe D . Incorporation of 9-β-D-arabinofuranosyl-2-fluoroadenine into HL-60 cellular RNA and DNA Biochem Pharmacol 1986 35: 247–252

    CAS  PubMed  Google Scholar 

  31. Genini D, Budihardjo I, Plunkett W, Wang X, Carrera CJ, Cottam HB, Carson DA, Leoni LM . Nucleotide requirements for the in vitro activation of the apoptosis protein-activating factor-1-mediated caspase pathway J Biol Chem 2000 275: 29–34

    CAS  PubMed  Google Scholar 

  32. Silber R, Degar B, Costin D, Newcomb EW, Mani M, Rosenberg CR, Morse L, Drygas JC, Canellakis ZN, Potmesil M . Chemosensitivity of lymphocytes from patients with B-cell chronic lymphocytic leukemia to chlorambucil, fludarabine, and camptothecin analogs Blood 1994 84: 3440–3446

    CAS  PubMed  Google Scholar 

  33. Thomas A, El Rouby S, Reed JC, Krajewski S, Silber R, Potmesil M, Newcomb EW . Drug-induced apoptosis in B-cell chronic lymphocytic leukemia: relationship between p53 gene mutation and bcl-2/bax proteins in drug resistance Oncogene 1996 12: 1055–1062

    CAS  PubMed  Google Scholar 

  34. Saven A, Piro LD . 2-Chlorodeoxyadenosine: a newer purine analog active in the treatment of indolent lymphoid malignancies Ann Intern Med 1994 120: 784–791

    CAS  PubMed  Google Scholar 

  35. Griffig J, Koob R, Blakley RL . Mechanisms of inhibition of DNA synthesis by 2-chlorodeoxyadenosine in human lymphoblastic cells Cancer Res 1989 49: 6923–6928

    CAS  PubMed  Google Scholar 

  36. Hentosh P, Koob R, Blakley RL . Incorporation of 2-halogeno-2′-deoxyadenosine 5-triphosphates into DNA during replication by human polymerases alpha and beta J Biol Chem 1990 265: 4033–4040

    CAS  PubMed  Google Scholar 

  37. Hentosh P, Tibudan M . In vitro transcription of DNA containing 2-chloro-2′-deoxyadenosine monophosphate Mol Pharmacol 1995 48: 897–904

    CAS  PubMed  Google Scholar 

  38. Bartik MM, Kay NE . Inhibition of IL-2 production mediates fludarabine-induced suppression of normal human T cell activation leading to apoptosis Proc Am Assoc Cancer Res 1998 39: 469

    Google Scholar 

  39. Collins RJ, Harmon BV, Souvlis T, Pope JH, Kerr JF . Effect of cycloheximide on B-chronic lymphocytic leukemic and normal lymphocytes in vitro: induction of apoptosis Br J Cancer 1991 64: 518–522

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Wright JC, Dolgopol VB, Logan M, Prigot A, Wright L . Clinical evaluation of puromycin in human neoplastic disease AMA Arch Intern Med 1955 115: 61–77

    Google Scholar 

  41. Romano MF, Lamberti A, Tassone P, Alfinito F, Costantini S, Chiurazzi F, Defrance T, Bonelli P, Tuccillo F, Turco MC, Venuta S . Triggering of CD40 antigen inhibits fludarabine-induced apoptosis in B chronic lymphocytic leukemia cells Blood 1998 92: 990–995

    CAS  PubMed  Google Scholar 

  42. Kitada S, Zapata JM, Andreeff M, Reed JC . Bryostatin and CD40-ligand enhance apoptosis resistance and induce expression of cell survival genes in B-cell chronic lymphocytic leukaemia Br J Haematol 1999 106: 995–1004

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported in part by grants CA32839, CA81534, CA77339 and Cancer Center Support Grant P30 CA16672 from the National Cancer Institute. PH is the recipient of a Physicians Referral Service Research Award from the University of Texas MD Anderson Cancer Center. EVDN is the recipient of a grant from the Commission du Patrimoine, Catholic University of Louvain, Belgian Hematological Society, and from the Commission for Educational Exchange, Brussels, Belgium.

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Authors and Affiliations

  1. Department of Experimental Therapeutics Leukemia, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA

    P Huang, A Sandoval, E Van Den Neste & W Plunkett

  2. Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA

    MJ Keating

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  1. P Huang
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  2. A Sandoval
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  3. E Van Den Neste
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  4. MJ Keating
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Huang, P., Sandoval, A., Van Den Neste, E. et al. Inhibition of RNA transcription: a biochemical mechanism of action against chronic lymphocytic leukemia cells by fludarabine. Leukemia 14, 1405–1413 (2000). https://doi.org/10.1038/sj.leu.2401845

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