Establishment and characterization of a megakaryoblast cell line with amplification of MLL.
A new cell line with megakaryoblastic features, designated UoC-M1, was established from the malignant cells of a 68-year-old patient with acute myeloid leukemia. The patient's leukemic cells reacted with alpha-naphthyl acetate esterase and acid phosphatase and expressed CD7, CD24, CD34, CD38, CD45, HLA-DR and CD61. Cytogenetic analysis of the patient's malignant cells (and of the UoC-M1 cells) showed a human, male hypodiploid karyotype with many chromosome rearrangements and marker chromosomes. Spectral karyotyping (SKY) analysis complemented the G-banded karyotyping and clarified several chromosomal translocations and identified the marker chromosomes. Fluorescence in situ hybridization (FISH) and SKY analysis demonstrated that one marker chromosome contained three segments of chromosome 9 interspersed with three segments of chromosome 11, as well as a portion of chromosome 19. FISH analysis with a probe for MLL revealed that the UoC-M1 cells contained four copies of the MLL gene. Southern blot analysis determined that the MLL gene had a germline profile while Northern and Western analyses showed that the MLL mRNAs and protein were of the appropriate sizes. This is the first report of amplification of the MLL gene which may be an additional mechanism of leukemogenesis or disease progression.
Allen
R J
RJ
Department of Pediatrics, University of Chicago, IL, USA.
Smith
S D
SD
Moldwin
R L
RL
Lu
M M
MM
Giordano
L
L
Vignon
C
C
Suto
Y
Y
Harden
A
A
Tomek
R
R
Veldman
T
T
Ried
T
T
Larson
R A
RA
Le Beau
M M
MM
Rowley
J D
JD
Zeleznik-Le
N
N
D000368:Aged; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000235:genetics*; D005784:Gene Amplification; D015321:Gene Rearrangement; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D016130:Immunophenotyping; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000473:pathology*; D008297:Male; D008533:Megakaryocytes / Q000166:cytology* / Q000502:physiology; D008854:Microscopy, Electron; D051788:Myeloid-Lymphoid Leukemia Protein; D011519:Proto-Oncogenes; D014157:Transcription Factors; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; C497422:KMT2A protein, human; D014157:Transcription Factors; D051788:Myeloid-Lymphoid Leukemia Protein; D011495:Histone-Lysine N-Methyltransferase