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Molecular Targeted Regulation Studies and Therapy

Targeted down-regulation of MLL-AF9 with antisense oligodeoxyribonucleotide reduces the expression of the HOXA7and -A10 genes and induces apoptosis in a human leukemia cell line, THP-1

Abstract

The MLL gene is frequently rearranged in leukemias, and MLL chimeric proteins generated by chromosomal translocations play crucial roles in leukemogenesis. Targets of murine Mll include HOX proteins that regulate body pattern formation and hematopoiesis. However, it is not known whether or not the MLL chimeric proteins regulate the HOX gene expression in human leukemia. To address this issue, THP-1 cells, a human leukemia cell line expressing MLL-AF9, were treated with antisense oligodeoxyribonucleotide (ODN) complementary to the coding sequence of the MLL-AF9 junction. Down-regulation of the MLL-AF9 transcript was accompanied by the reduced expression of the HOXA7 and -A10 genes, but not of the HOXA2, -A4, -A5, and -A9 genes. The number of viable cells cultured with 20 μM antisense ODN for 5 days was 10-fold lower than that of the sense ODN-treated control. And the number of the annexin V/propidium iodide apoptotic cells in the antisense ODN-treated cells after 3 days of culture was two-fold higher than that in the control. Staining of the antisense ODN-treated cells with Hoechst 33258 showed the morphology characteristic to apoptosis. These results indicate that MLL-AF9 regulates the expression of the selected HOX genes as well as prevents the leukemic cells from apoptosis.

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References

  1. Cimino G, Moir DT, Canaani O, Williams K, Crist WM, Katzav S, Cannizzaro L, Lange B, Nowell PC, Croce CM, Canaani E . Cloning of ALL-1, the locus involved in leukemias with the t(4;11)(q21;q23), t(9;11)(p22;q23), and t(11;19)(q23;p13) chromosome translocations Cancer Res 1991 51: 6712–6714

    CAS  PubMed  Google Scholar 

  2. Ziemin-van der Poel S, McCabe NR, Gill HJ, Espinosa R, Patel Y, Harden A, Rubinelli P, Smith SD, LeBeau MM, Rowley JD, Diaz MO . Identification of a gene, MLL, that spans the breakpoint in 11q23 translocations associated with human leukemias Proc Natl Acad Sci USA 1991 88: 10735–10739

    Article  CAS  Google Scholar 

  3. Gu Y, Nakamura T, Alder H, Prasad R, Canaani O, Cimino G, Croce CM, Canaani E . The t(4;11) chromosome translocation of human acute leukemias fuses the ALL-1 gene, related to Drosophila trithorax, to the AF-4 gene Cell 1992 71: 701–708

    Article  CAS  Google Scholar 

  4. Tkachuk DC, Kohler S, Cleary ML . Involvement of a homologue of Drosophila Trithorax by 11q23 chromosomal translocations in acute leukemias Cell 1992 71: 691–700

    Article  CAS  Google Scholar 

  5. Biondi A, Cimino G, Pieters R, Pui CH . Biological and therapeutic aspects of infant leukemia Blood 2000 96: 24–33

    CAS  Google Scholar 

  6. DiMartino JF, Cleary ML . MLL Rearrangements in haematological malignancies: lessons from clinical and biological studies Br J Haematol 1999 106: 614–626

    Article  CAS  Google Scholar 

  7. Pui CH, Relling MV, Rivera GK, Hancock ML, Raimondi SC, Heslop HE, Santana VM, Ribeiro RC, Sandlund JT, Mahmoud HH, Evans WE, Crist WM, Krance RA . Epipodophyllotoxin-related acute myeloid leukemia: a study of 35 cases Leukemia 1995 9: 1990–1996

    CAS  PubMed  Google Scholar 

  8. Zeleznik-Le NJ, Harden AM, Rowley JD . 11q23 translocations split the ‘AT-hook’ cruciform DNA-binding region and the transcriptional repression domain from the activation domain of the mixed-lineage leukemia (MLL) gene Proc Natl Acad Sci USA 1994 91: 10610–10614

    Article  CAS  Google Scholar 

  9. Arakawa H, Nakamura T, Zhadanov AB, Fidanza V, Yano T, Bullrich F, Shimizu M, Blechman J, Mazo A, Canaani E, Croce CM . Identification and characterization of the ARP1 gene, a target for the human acute leukemia ALL1 gene Proc Natl Acad Sci USA 1998 95: 4573–4578

    Article  CAS  Google Scholar 

  10. Rozenblatt-Rosen O, Rozovskaia T, Bukarov D, Sedkov Y, Tillib S, Blechman J, Nakamura T, Croce CM, Mazo A, Canaani E . The C-terminal SET domains of ALL1 and TRITHORAX interact with the INI1 and SNR1 proteins, components of the SWI/SNF complex Proc Natl Acad Sci USA 1998 95: 4152–4157

    Article  CAS  Google Scholar 

  11. Morrissey JJ, Raney S, Cleary ML . The FEL (AF-4) protein donates transcriptional activation sequences to HRX-FEL fusion proteins in leukemias containing t(4;11)(q21;q23) chromosomal translocations Leukemia Res 1997 21: 911–917

    Article  CAS  Google Scholar 

  12. Prasad R, Yano T, Sorio C, Nakamura T, Rallapalli R, Gu Y, Leshkowitz D, Croce CM, Canaani E . Domains with transcriptional regulatory activity within the ALL1 and AF4 proteins involved in acute leukemia Proc Natl Acad Sci USA 1995 92: 12160–12164

    Article  CAS  Google Scholar 

  13. Rubnitz JE, Morrissey J, Savage PA, Cleary ML . ENL, the gene fused with HRX in t(11;19) leukemias, encodes a nuclear protein with transcriptional activation potential in lymphoid and myeloid cells Blood 1994 84: 1747–1752

    CAS  Google Scholar 

  14. Corral J, Lavenir I, Impey L, Warren AJ, Forster A, Larson TA, Bell S, McKenzie ANJ, King G, Rabbitts TH . An Mll-Af9 fusion gene made by homologous recombination causes acute leukemia in chimeric mice: a method to create fusion oncogenes Cell 1996 85: 853–861

    Article  CAS  Google Scholar 

  15. Dobson CL, Warren AJ, Pannell R, Forster A, Rabbitts TH . Tumorigenesis in mice with a fusion of the leukaemia oncogene Mll and the bacterial lacZ gene EMBO J 2000 19: 843–851

    Article  CAS  Google Scholar 

  16. Chaplin T, Ayton P, Bernard OA, Saha V, Della Valle V, Hillion J, Gregorini A, Lillington D, Berger R, Young BD . A novel class of zinc finger/leucine zipper genes identified from the molecular cloning of the t(10;11) translocation in acute leukemia Blood 1995 85: 1435–1441

    CAS  Google Scholar 

  17. Prasad R, Lesshkowitz D, Gu Y, Alder H, Nakamura T, Saito H, Huebner K, Berger R, Croce CM, Canaani E . Leucine-zipper dimerization motif encoded by the AF17 gene fused to ALL-1 (MLL) in acute leukemia Proc Natl Acad Sci USA 1994 91: 8107–8111

    Article  CAS  Google Scholar 

  18. Yu BD, Hess JL, Horning SE, Brown GAJ, Korsmeyer SJ . Altered Hox expression and segmental identity in Mll-mutant mice Nature 1995 378: 505–508

    Article  CAS  Google Scholar 

  19. Gehring WJ . Homeoboxes in the study of development Science 1987 236: 1245–1252

    Article  CAS  Google Scholar 

  20. Kessel M, Gruss P . Murine developmental control genes Science 1990 249: 374–379

    Article  CAS  Google Scholar 

  21. Graham A, Papalopulu N, Krumlauf R . The murine and Drosophila homeobox genes complexes have common features of organization and expression Cell 1989 57: 367–378

    Article  CAS  Google Scholar 

  22. Scott MP . Vertebrate homeobox gene nomenclature Cell 1992 71: 551–553

    Article  CAS  Google Scholar 

  23. Lawrence HJ, Sauvageau G, Humphries RK, Largman C . The role of homeobox genes in normal and leukemic hematopoiesis Stem Cells 1996 14: 281–291

    Article  CAS  Google Scholar 

  24. van Oostveen JW, Bijl JJ, Raaphorst FM, Walboomers JJM, Meijer CJLM . The role of Homeobox genes in normal hematopoiesis and hematological malignancies Leukemia 1999 13: 1675–1690

    Article  CAS  Google Scholar 

  25. Sauvageau G, Lansdorp PM, Eaves CJ, Hogge DE, Dragowska WH, Reid DS, Largman C, Lawrence HJ, Humphries RK . Differential expression of homeobox genes in functionally distinct CD34+ subpopulations of human bone marrow cells Proc Natl Acad Sci USA 1994 91: 12223–12227

    Article  CAS  Google Scholar 

  26. Thorsteinsdottir U, Sauvageau G, Hough MR, Dragowska WH, Lansdorp PM, Lawrence HJ, Largman C, Humphries RK . Overexpression of HOXA10 in murine hematopoietic cells perturbs both myeloid and lymphoid differentiation and leads to acute myeloid leukemia Mol Cell Biol 1997 17: 495–505

    Article  CAS  Google Scholar 

  27. Kroon E, Krosl J, Thorsteinsdottir U, Baban S, Buchberg AM, Sauvageau G . Hoxa9 transforms primary bone marrow cells through specific collaboration with Meis 1a but not Pbx 1b EMBO J 1998 17: 3714–3725

    Article  CAS  Google Scholar 

  28. Tsuchiya S, Yamabe M, Yamaguchi Y, Kobayashi Y, Konno T, Tada K . Establishment and characterization of a human acute monocytic leukemia cell line (THP-1) Int J Cancer 1980 26: 171–176

    Article  CAS  Google Scholar 

  29. Lozzio CB, Lozzio BB . Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome Blood 1975 45: 321–334

    CAS  PubMed  Google Scholar 

  30. Gallagher RE, Collins SJ, Trujillo J, McCredie K, Ahearn M, Tsai S, Metzgar R, Aulakh G, Ting R, Ruscetti F, Gallo RC . Characterization of the continuous differentiating myeloid cell line (HL-60) from a patient with acute promyelocytic leukemia Blood 1979 54: 713–733

    CAS  PubMed  Google Scholar 

  31. Brady G, Barbara M, Iscove NN . Representative in vitro cDNA amplification from individual hematopoietic cells and colonies Meth Mol Cell Biol 1990 2: 17–25

    CAS  Google Scholar 

  32. Kawagoe H, Humphries RK, Blair A, Sutherland HJ, Hogge DE . Expression of HOX genes, HOX-cofactors, and MLL in phenotypically and functionally defined subpopulations of leukemic and normal human hematopoietic cells Leukemia 1999 13: 687–698

    Article  CAS  Google Scholar 

  33. Vieille-Grosjean I, Roullot V, Courtois G . Lineage and stage specific expression of HOX 1 genes in the human hematopoietic system Biochem Biophys Res Commun 1992 183: 1124–1130

    Article  CAS  Google Scholar 

  34. Vermes I, Haanen C, Reutelingsperger CPM . A novel assay for apoptosis: flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labeled annexin V J Immunol Meth 1995 180: 39–52

    Article  Google Scholar 

  35. Schreiner SA, García-Cuéllar, MP, Fey GH, Slany RK . The leukemogenic fusion of MLL with ENL creates a novel transcriptional transactivator Leukemia 1999 13: 1525–1533

    Article  CAS  Google Scholar 

  36. Dupé V, Ghyselinck NB, Thomazy V, Nagy L, Davies PJA, Chambon P, Mark M . Essential roles of retinoic acid signaling in interdigital apoptosis and control of BMP-7 expression in mouse autopods Dev Biol 1999 208: 30–43

    Article  Google Scholar 

  37. Nakamura T, Largaespada DA, Shaughnessy JD Jr, Jenkins NA, Copeland NG . Cooperative activation of Hoxa and Pbx1-related genes in murine myeloid leukemias Nat Genet 1996 12: 149–153

    Article  CAS  Google Scholar 

  38. Lawrence HJ, Sauvageau G, Ahmadi N, Lopez AR, LeBeau MM, Link M, Humphries K, Largman C . Stage- and lineage-specific expression of the HOXA10 homeobox gene in normal and leukemic hematopoietic cells Exp Hematol 1995 23: 1160–1166

    CAS  PubMed  Google Scholar 

  39. Akao Y, Mizoguchi H, Misiura K, Stec WJ, Seto M, Ohishi N, Yagi K . Antisense oligodeoxyribonucleotide against the MLL-LTG19 chimeric transcript inhibits cell growth and induces apoptosis in cells of an infantile leukemia cell line carrying the t(11;19) chromosomal translocation Cancer Res 1998 58: 3773–3776

    CAS  PubMed  Google Scholar 

  40. McGahon A, Bissonnentte R, Schmitt M, Cotter KM, Green DR, Cotter TG . BCR-ABL maintains resistance of chronic myelogeneous leukemia cells to apoptotic cell death Blood 1994 83: 1179–1187

    CAS  Google Scholar 

  41. Inaba T, Inukai T, Yoshihara T, Seyschab H, Ashmun RA, Canman CE, Laken SJ, Kastan MB, Look T . Reversal of apoptosis by the leukaemia-associated E2A-HLF chimaeric transcription factor Nature 1996 382: 541–544

    Article  CAS  Google Scholar 

  42. Dobson CL, Warren AJ, Pannell R, Forster A, Lavenir I, Corral J, Smith AJH, Rabbitts TH . The Mll-AF9 gene fusion in mice controls myeloproliferation and specifies acute myeloid leukaemogenesis EMBO J 1999 18: 3564–3574

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by a Grant for Priority Area and a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (to KS).

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Kawagoe, H., Kawagoe, R. & Sano, K. Targeted down-regulation of MLL-AF9 with antisense oligodeoxyribonucleotide reduces the expression of the HOXA7and -A10 genes and induces apoptosis in a human leukemia cell line, THP-1. Leukemia 15, 1743–1749 (2001). https://doi.org/10.1038/sj.leu.2402262

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