Summary
The presence of activated transforming genes was investigated in four patients with therapy-related leukemia and in three with therapy-related myelodysplastic syndrome. DNA of bone marrow cells from six of the patients exhibited transforming activity in the tumorigenicity assay. Five of the six patients who were positive in the tumorigenicity assay contained activated N-ras oncogenes, and three contained activated K-ras oncogenes. Thus, concurrent activation of N-ras and K-ras oncogenes was observed in two patients.
In vitro DNA amplification followed by oligonucleotide dot-blot analysis was used to investigate mutations in codons 12, 13, and 61 of the N-ras and K-ras oncogenes. Two patients exhibited an N-ras mutation, substituting aspartic acid (GAT) for glycine (GGT), and three patients exhibited an N-ras codon 13 mutation, substituting valine (GTT) for glycine. Two patients exhibited K-ras codon 12 mutations, substituting aspartic acid (GAT) or cysteine (TGT) for glycine (GGT), respectively, and one case exhibited a K-ras codon 61 mutation, substituting lysine (AAA) for glutamic acid (CAA). Cytogenetic analysis revealed that loss of chromosome 7 was frequent (four patients: 57%). Our data indicate that activation of N-ras and K-ras genes, as well as loss of heterozygosity for specific alleles on chromosome 7, plays a more important role in the leukemogenesis of both therapy-related leukemia and myelodysplastic syndrome.
Similar content being viewed by others
References
Anderson RL, Bagby GC, Richert-Boe K, Magenis RE, Koler RD (1981) Therapy-related preleukemic syndrome. Cancer 47: 1867–1871
Bar-Eli M, Ahuja H, Gonzalez-Cadavid N, Foti A, Cline MJ (1989) Analysis of N-ras exon-1 mutations in myelodysplastic syndromes by polymerase chain reaction and direct sequencing. Blood 73: 281–283
Bartram CR, Ludwig W, Hiddemann W, Lyons J, Buschle M, Ritter J, Harbott J, Frohlich A, Janssen JWG (1989) Acute myeloid leukemia: analysis ofras gene mutations and clonality defined by polymorphic X-linked loci. Leukemia 3: 247–256
Benitez J, Carbonell F, Ferro T, Prieto F, Fayos JS (1986) Cytogenetic studies in 18 patients with secondary blood disorders. Cancer Genet Cytogenet 22: 309–317
Bos JL, Toksoz D, Marshall CJ, Verlaan-de Vries M, Veeneman GH, Vander EbAJ, Van Boom JH, Janssen JWG, Steenvoorden ACM (1985) Amino acid substitutions at codon 13 of the N-ras oncogene in human acute myeloid leukemia. Nature 315: 726–730
Bos JL, Vries MV, van der Eb AJ, Janssen JWG, Delwel R, Lowenberg B, Colly LP (1987) Mutations in N-ras predominate in acute myeloid leukemia. Blood 69: 1237–1241
Coltman C (1982) Treatment-related leukemia. In: Bloomfield CD (ed) Acute leukemia. Martinus Nijhoff, The Hague, p 61
Diamond LE, Guerrero I, Pellicer A (1988) Concomitant K- and N-ras gene point mutations in clonal murine lymphoma. Mol Cell Biol 8: 2233–2236
Farr CJ, Saiki RK, Erlich HA, McCormick F, Marshall CJ (1988) Analysis of RAS gene mutations in acute myeloid leukemia by polymerase chain reaction and oligonucleotide probes. Proc Natl Acad Sci USA 85: 1629–1633
Fasano O, Birnbaum D, Edlund L, Fogh J, Wigler M (1984) New human transforming genes detected by a tumorigenicity assay. Mol Cell Biol 4: 1695–1705
Graham JF, van der Eb AJ (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52: 456–467
Hirai H, Tanaka S, Azuma M, Anraku Y, Kobayashi Y, Fujisaria M, Okabe T, Urabe T, Takaku F (1985) Transforming genes in human leukemia cells. Blood 66: 1371–1378
Hirai H, Kobayashi Y, Mano H, Hagiwara K, Maru Y, Omine M, Mizoguchi H, Nishizima J, Takaku F (1987) A point mutation at codon 13 of the N-ras oncogene in myelodysplastic syndrome. Nature 327: 430–432
Inokuchi K, Tajika K, Ito T, Ogata K, Yamada T, Tanabe Y, Ohki I, Dan K, Kuriya S, Nomura T, Shinohara T (1988) Secondary myelodysplastic syndrome with monosomy 7. Jpn J Clin Hematol 29: 718–722
Inokuchi K, Komiya I, Dan K, Kuriya S, Shinohara T, Nomura T (1990) TdT-positive, SmIg-negative B precursor cell leukemia with Burkitt morphology: a case report. Leuk Lymph 2: 251–255
Janssen JWG, Lyons J, Steenvoorden ACM, Seliger H, Bartram CR (1987) Concurrent mutations in two differentras genes in acute myelocytic leukemias. Nucleic Acid Res 15: 5669–5680
Janssen JWG, Steenvorrden ACM, Lyons J, Anger B, Bohlke JU, Bos JL, Seliger H, Bartram CR (1987) RAS gene mutations in acute and chronic myelocytic leukemias, chronic myeloproliferative disorders and myelodysplastic syndromes. Proc Natl Acad Sci USA 84: 9228–9232
Kantarjian HM, Keating MJ, Walters RS, Smith TL, Cork A, McCredie K, Freirech EJ (1986) Therapy-related leukemia and myelodysplastic syndrome: clinical, cytogenetic, and prognostic features. J Clin Oncol 4: 1748–1757
Koeffler P, Rowley J (1985) Therapy-related acute non-lymphocytic leukemia. In: Wiernik P, Canellos G, Kyle R et al. (eds) Neoplastic Diseases of the blood. Churchill Livingstone, New York, p 357
LeBeau MM, Albain KS, Larson RA, Vardiman JW, Davis EM, Blough RR, Golomb HM, Rowley JD (1986) Clinical and cytogenetic correlations in 63 patients with therapy-related myelodysplastic syndromes and acute nonlymphocytic leukemia: further evidence for characteristic abnormalities of chromosomes no. 5 and 7. J Clin Oncol 4: 325–345
Liu E, Hjelle B, Bishop M (1988) Transforming genes in chronic myelogenous leukemia. Proc Natl Acad Sci USA 85: 1952–1956
Mano H, Ishikawa F, Hirai H, Takaku F (1989): Mutations of N-ras oncogene in myelodysplastic syndromes and leukemias detected by polymerase chain reaction. Jpn J Cancer Res 80: 102–106
Michels SD, McKenna RW, Arthur DC, Brunning RD (1985) Therapy-related acute myeloid leukemia and myelodysplastic syndrome: a clinical and morphologic study of 65 cases. Blood 65: 1364–1372
Neri A, Knowles DM, Greco A, McCormick F, Dalla-Favera R (1988) Analysis of RAS oncogene mutations in human lymphoid malignancies. Proc Natl Acad Sci USA 85: 9268–9272
Ogata K, Futaki M, Inokuchi K, Gomi S, Ohki I, Kuwabara T, Dan K, Kuriya S, Nomura T, Shinohara T (1989) Successful treatment of secondary erythroleukemia with androgen. Jpn J Clin Hematol 30: 1079–1083
Pedersen-Bjergaard J, Janssen WG, Lyons J, Philip P, Bartram CR (1988) Point mutation of theras proto-oncogenes and chromosome aberrations in acute nonlymphocytic leukemia and preleukemia related to therapy with alkylating agents. Cancer Res 48: 1812–1817
Pedersen-Bjergaard J, Larsen SO (1982) Incidence of acute nonlymphocytic leukemia, and acute myeloproliferative syndrome up to 10 years after treatment of Hodgkin's disease. N Engl J Med 307: 965–971
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487–491
Shimada T, Inokuchi K, Nienhuis AW (1987) Site-specific demethylation and normal chromatin structure of the human dihydroforate reductase gene promoter after transfection into CHO cells. Mol Cell Biol 7: 2830–2837
Vries MV, Bogaard ME, van den Elst H, van Boom J, van der Eb AJ, Bos JL (1986) A dot-blot screening procedure for mutatedras oncogenes using synthetic oligo-deoxynucleotides. Gene 50: 313–320
Author information
Authors and Affiliations
Additional information
This work was supported in part a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan
Rights and permissions
About this article
Cite this article
Inokuchi, K., Amuro, N., Futaki, M. et al. Transforming genes and chromosome aberrations in therapy-related leukemia and myelodysplastic syndrome. Ann Hematol 62, 211–216 (1991). https://doi.org/10.1007/BF01729834
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01729834