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Ratio of bcl-xshort to bcl-xlong Is Different in Good- and Poor-Prognosis Subsets of Acute Myeloid Leukemia

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Abstract

Background

Acute myeloid leukemia (AML) is a heterogeneous collection of leukemic disorders ranging from chemotherapy-sensitive subsets [inversion 16 and t(8;21)], which often can be cured with cytosine arabinoside alone, to the most resistant subsets, which can survive even supralethal levels of combination alkylator chemotherapy (cytogenetic subsets monosomy 5 and monosomy 7).

Materials and Methods

To analyze the expression of BCL-2 family genes, which are expressed in these subsets of AML, we used PCR sequence amplification reactions that are dependent on oligonucleotide primers representing the BH1 and BH2 homology domains to generate the unique regions between BH1 and BH2. These primers are conserved among all members of the BCL-2 gene family and are separated by a 150 nucleotide region sequence between the BH1 and BH2 domains. The PCR products unique to each BCL-2 family member were cloned directionally into sequencing vectors. The identity of the insert of each clone was determined by slot-blots of the DNA amplified from individual colonies and by hybridization with radioactive probes specific to the bcl-2, bcl-x, or bax genes.

Results

We found that bcl-2 is the predominant member expressed in AML samples with a poor prognosis (−5, −7), whereas the transcripts of bcl-x are higher than those of bcl-2 in the AML samples with a good prognosis [invl6, t(8;21)]. No significant difference in bax expression was detected between AML subsets of good and bad prognosis. The ratio of bcl-xlong, which inhibits apoptosis, to bcl-xshort, which promotes apopto-sis, was determined by amplification with a pair of primers specific to bcl-x followed by separation of the PCR product on agarose gels. Bcl-xlong and bcl-xshort appeared as bands of different molecular mass on a molecular weight gel and were visualized by ethidium bromide staining or Southern blot analysis with a bcl-x-specific probe.

Conclusions

We found that the ratio of bcl-x long to bcl-x short was higher in the AML patients with a poor prognosis. These experiments showed that the levels of BCL-2 family members in the leukemia cells of good- and poor-prognosis subsets are different. In addition, novel members of the BCL-2 family were isolated from the cells of AML patients of either prognosis.

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References

  1. Marlton P, Keating M, Kantarjian H, Pierce S, O’Brien S, Freireich EJ, Estey E. (1995) Cytogenetic and clinical correlates in AML patients with abnormalities of chromosome 16. Leukemia 9: 965–971.

    PubMed  CAS  Google Scholar 

  2. Walker H, Smith FJ, Betts DR. (1994) Cytogenetics in acute myeloid leukemia. Blood Rev. 8: 30–36.

    Article  CAS  PubMed  Google Scholar 

  3. Maseki N, Miyoshi H, Shimizu K, Homma C, Ohki M, Sakurai M, Kaneko Y. (1993) The 8;21 chromosome translocation in acute myeloid leukemia is always detectable by molecular analysis using AML1. Blood 81: 1573–1579.

    PubMed  CAS  Google Scholar 

  4. Symes PH, Williams ME, Flessa HC, Srivastava AK, Swerdlow SH. (1993) Acute promyelocytic leukemia with the pseudo-Chediak-Higashi anomaly and molecular documentation of t(15; 17) chromosomal translocation. Am. J. Clin. Pathol. 99: 622–627.

    Article  CAS  PubMed  Google Scholar 

  5. Hoffmann L, Moller P, Pedersen BJ, Waage A, Pedersen M, Hirsch FR. (1995) Therapy-related acute promyelocytic leukemia with t(15;17)(q22; q12) following chemotherapy with drugs targeting DNA topoisomerase II. A report of two cases and a review of the literature. Ann. Oncol. 6: 781–788.

    Article  CAS  PubMed  Google Scholar 

  6. Baranger L, Baruchel A, Leverger G, Schaison G, Berger R. (1990) Monosomy-7 in childhood hemopoietic disorders. Leukemia 4: 345–349.

    PubMed  CAS  Google Scholar 

  7. Stephenson J, Lizhen H, Mufti GJ. (1995) Possible co-existence of RAS activation and monosomy 7 in the leukemic transformation of myelodysplastic syndromes. Leuk. Res. 19: 741–748.

    Article  CAS  PubMed  Google Scholar 

  8. Vogler WR. (1992) Strategies in the treatment of acute myelogenous leukemia. Leuk. Res. 16: 1143–1148.

    Article  CAS  PubMed  Google Scholar 

  9. Downing JR, Head DR, Curcio BA, et al. (1993) An AML1/ETO fusion transcript is consistently detected by RNA-based polymerase chain reaction in acute myelogenous leukemia containing the (8;21)(q22;q22) translocation. Blood 81:2860–2865.

    PubMed  CAS  Google Scholar 

  10. Lo CF, Diverio D, D’Adamo F, Avvisati G, Alimena G, Nanni M, Alcalay M, Pandolfi PP, Pelicci PG. (1992) PML/RAR-alpha rearrangement in acute promyelocytic leukemias apparently lacking the t(15;17) translocation. Eur. J. Haematol. 48: 173–177.

    Google Scholar 

  11. Nucifora G, Dickstein JI, Torbenson V, Roulston D, Rowley JD, Vardiman JW. (1994) Correlation between cell morphology and expression of the AML1/ETO chimeric transcript in patients with acute myeloid leukemia without the t(8;21). Leukemia 8: 1533–1538.

    PubMed  CAS  Google Scholar 

  12. de Greef G, Hagemeijer A, Morgan R, Wijsman J, Hoefsloot LH, Sandberg AA, Sacchi N. (1995) Identical fusion transcript associated with different breakpoints in the AML1 gene in simple and variant t(8;21) acute myeloid leukemia. Leukemia 9: 282–288.

    PubMed  Google Scholar 

  13. Reed JC. (1995) Bcl-2 family proteins: regulators of chemoresistance in cancer. Toxicol. Lett. 155: 82–84.

    Google Scholar 

  14. Korsmeyer SJ. (1995) Regulators of cell death. Trends Genet. 11: 101–105.

    Article  CAS  PubMed  Google Scholar 

  15. Craig RW. (1995) The bcl-2 gene family. Semin. Cancer Biol. 6: 35–43.

    Article  CAS  PubMed  Google Scholar 

  16. Yin XM, Oltval ZN, Korsmeyer SJ. (1994) BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax [see comments]. Nature 369: 321–323.

    Article  CAS  PubMed  Google Scholar 

  17. Boise LH, Gonzalez GM, Postema CE, et al. (1993) Bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell 74: 597–608.

    Article  CAS  PubMed  Google Scholar 

  18. Kozopas KM, Yang T, Buchan HL, Zhou P, Craig RW. (1993) MCL1, a gene expressed in programmed myeloid cell differentiation, has sequence similarity to BCL2. Proc. Natl. Acad. Sci. USA 90: 3516–3520.

    Article  CAS  PubMed  Google Scholar 

  19. Miyashita T, Reed JC. (1995) Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80: 293–299.

    Article  CAS  PubMed  Google Scholar 

  20. Park JR, Bernstein ID, Hockenbery DM. (1995) Primitive human hematopoietic precursors express Bcl-x but not Bcl-2. Blood 86: 868–876.

    PubMed  CAS  Google Scholar 

  21. Chittenden T, Harrington EA, O’Connor R, Flemington C, Lutz RJ, Evan GI, Guild BC. (1995) Induction of apoptosis by the Bcl-2 homologue Bak. Nature 374: 733–736.

    Article  CAS  PubMed  Google Scholar 

  22. Farrow SN, White JH, Martinou I, et al. (1995) Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19K. Nature 374:731–733.

    Article  CAS  PubMed  Google Scholar 

  23. Karsan A, Yee E, Kaushansky K, Harlan JM. (1996) Cloning of human bcl-2 homologue: Inflammatory cytokines induce human Al in cultured endothelial cells. Blood 87: 3089–3096.

    PubMed  CAS  Google Scholar 

  24. Kiefer MC, Brauer MJ, Powers VC, Wu JJ, Umansky SR, Tomei LD, Barr PJ. (1995) Modulation of apoptosis by the widely distributed Bcl-2 homologue Bak. Nature 374: 736–739.

    Article  CAS  PubMed  Google Scholar 

  25. Lin EY, Orlofsky A, Berger MS, Prystowsky MB. (1993) Characterization of Al, a novel hemopoi-etic-specific early-response gene with sequence similarity to bcl-2. J. Immunol 151: 1979–1988.

    PubMed  CAS  Google Scholar 

  26. Yang E, Zha J, Jockel J, Boise LH, Thompson CB, Korsmeyer SJ. (1995) Bad, a heterodimeric partner for Bcl-XL and Bcl-2, displaces Bax and promotes cell death. Cell 80: 285–291.

    Article  CAS  PubMed  Google Scholar 

  27. Maung ZT, MacLean FR, Reid MM, Pearson AD, Proctor SJ, Hamilton PJ, Hall AG. (1994) The relationship between bcl-2 expression and response to chemotherapy in acute leukaemia. Br. J. Haematol. 88: 105–109.

    Article  CAS  PubMed  Google Scholar 

  28. Keith FJ, Bradbury DA, Zhu YM, Russell NH. (1995) Inhibition of bcl-2 with antisense oligonucleotides induces apoptosis and increases the sensitivity of AML blasts to ara-C. Leukemia 9: 131–138.

    PubMed  CAS  Google Scholar 

  29. Minn AJ, Rudin CM, Boise LH, Thompson C. (1995) Expression of bcl-xl can confer a multidrug resistance phenotype. Blood 86: 1903–1910.

    PubMed  CAS  Google Scholar 

  30. Dole MG, Jasty R, Cooper MJ, Thompson CB, Nunez G, Castle VP. (1995) Bcl-xl is expressed in neuroblastoma cells and modulates chemotherapy-induced apoptosis. Cancer Res. 55: 2576–2582.

    PubMed  CAS  Google Scholar 

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Acknowledgements

The authors acknowledge support from NCI P01 55164, from the George and Barbara Bush Leukemia Research Fund, the Anderson Chair for Cancer Treatment and Research at the University of Texas M.D. Anderson Cancer Center, the Ensign Professorship of Medicine at the Yale University School of Medicine, and the Hull Development Fund of the Yale Cancer Center.

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Authors and Affiliations

  1. University of Texas M. D. Anderson Cancer Center, Houston, Texas, USA

    G. Deng, C. Lane, S. Kornblau, A. Goodacre, V. Snell & M. Andreeff

  2. Section of Medical Oncology, WWW221, Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut, 06520-8032, USA

    Albert B. Deisseroth

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  1. G. Deng
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  2. C. Lane
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  3. S. Kornblau
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  4. A. Goodacre
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  5. V. Snell
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  6. M. Andreeff
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  7. Albert B. Deisseroth
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Correspondence to Albert B. Deisseroth.

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Communicated by E. Beutler.

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Deng, G., Lane, C., Kornblau, S. et al. Ratio of bcl-xshort to bcl-xlong Is Different in Good- and Poor-Prognosis Subsets of Acute Myeloid Leukemia. Mol Med 4, 158–164 (1998). https://doi.org/10.1007/BF03401913

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