Annals of Hematology

, Volume 87, Issue 12, pp 991–1002

The application of conventional cytogenetics, FISH, and RT-PCR to detect genetic changes in 70 children with ALL

  • Krystyna Soszynska
  • Barbara Mucha
  • Robert Debski
  • Katarzyna Skonieczka
  • Ewa Duszenko
  • Andrzej Koltan
  • Mariusz Wysocki
  • Olga Haus
Original Article

DOI: 10.1007/s00277-008-0540-6

Cite this article as:
Soszynska, K., Mucha, B., Debski, R. et al. Ann Hematol (2008) 87: 991. doi:10.1007/s00277-008-0540-6

Abstract

We investigated bone marrow cells of 70 acute lymphoblastic leukemia children by conventional cytogenetics (CC), fluorescence in situ hybridization (FISH), and reverse transcription polymerase chain reaction (RT-PCR) methods. CC and RT-PCR for fusion genes BCR/ABL, MLL/AF4, E2A/PBX1, TEL/AML1 were performed at diagnosis in each patient. FISH was performed to verify the presence of fusion genes and MLL rearrangements and to estimate the percentage of abnormal cells. Karyotypes were obtained in 59 (84%) of 70 cases. Thirty-five (59%) of 59 cases revealed chromosome aberrations. Hyperdiploidy >50 chromosomes was present in nine cases, hyperdiploidy 47–50 chromosomes in six, pseudodiploidy in 15, and hypodiploidy in five. BCR/ABL was present in two cases, PBX1/E2A in two, and TEL/AML1 in 14. MLL/AF4 was not found, but the rearrangements of MLL gene were present in five children. The addition of RT-PCR and FISH to CC was of the utmost importance. One of two Ph translocations and one of two t(1;19) were first revealed by RT-PCR. Moreover, FISH showed the percentage of TEL/AML1(+) cells that turned to be an important prognostic factor. The outcome was the best for the children with hyperdiploidy >50 chromosomes without structural changes. It was also good for those with TEL/AML1 present in ≥80% of cells without chromosome aberrations. The presence of pseudodiploidy correlated with poor outcome. The outcome for patients with t(9;22)–BCR/ABL or 11q23–MLL rearrangement was the worst in study group. The presence of BCR/ABL caused eight times increase of risk of death; MLL rearrangements caused 12 times increase.

Keywords

Acute lymphoblastic leukemiaChromosome aberrationsFISHRT-PCRPrognosis

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Krystyna Soszynska
    • 1
  • Barbara Mucha
    • 1
  • Robert Debski
    • 2
  • Katarzyna Skonieczka
    • 1
  • Ewa Duszenko
    • 3
  • Andrzej Koltan
    • 2
  • Mariusz Wysocki
    • 2
  • Olga Haus
    • 1
    • 3
  1. 1.Department of Clinical Genetics, Ludwik Rydygier Collegium MedicumNicolaus Copernicus UniversityBydgoszczPoland
  2. 2.Department of Pediatrics, Hematology and Oncology, Ludwik Rydygier Collegium MedicumNicolaus Copernicus UniversityBydgoszczPoland
  3. 3.Department of HematologyUniversity of MedicineWroclawPoland