Molecular characterization of 16p deletions associated with inversion 16 defines the critical fusion for leukemogenesis.
The inversion of chromosome 16 [inv(16)] in acute myeloid leukemia (AML) is associated with a p-arm deletion in a subset of patients. The inversion results in two fusion genes: 5'-CBFB/MYH11-3' on 16p and 5'-MYH11/CBFB-3' on 16q. We have studied cells from 42 patients with inv(16) (38 patients) or t(16;16) (four patients) to define the frequency and characteristics of the deletion further. Using fluorescence in situ hybridization (FISH) with probes from cosmids, cosmid contigs, and yeast artificial chromosomes (YACs), we found that six patients with inv(16) had a deletion of between 150 and 350 kb centromeric to the p-arm inversion breakpoint cluster region (p-ibc). This region was shown to contain the 5' portion of the myosin heavy chain (MYH11) gene. YACs containing the p-ibc, which had been useful as FISH probes in the diagnosis of inv(16), detected the inversion in deletion as well as nondeletion patient cells. Thus, the deleted region identified in patients is entirely contained within the human genomic content of the YACs. Southern blot experiments using probes flanking the p-ibc indicated that the deletion removes segments within 10 kb centromeric of the p-ibc. Reverse transcription-polymerase chain reaction (RT-PCR) using primers from the 5' region of CBFB and the 3' region of MYH11 (distal to the p-ibc) produced the 5'-CBFB/MYH11-3' chimeric transcript in inv(16)/del patients. These data confirm that the 5'-CBFB/MYH11-3' chimeric transcript, rather than the reciprocal 5'-MYH11/CBFB-3', is the critical product for chromosome 16-related leukemogenesis.
Department of Hematology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
P
P
Marlton
D F
DF
Claxton
P
P
Liu
E H
EH
Estey
M
M
Beran
M
M
LeBeau
J R
JR
Testa
F S
FS
Collins
J D
JD
Rowley
M J
MJ
Siciliano
D000208:Acute Disease; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D007446:Chromosome Inversion; D002874:Chromosome Mapping; D002885:Chromosomes, Human, Pair 16; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics*; D005091:Exons; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D007951:Leukemia, Myeloid / Q000145:classification / Q000235:genetics*; D008969:Molecular Sequence Data; D009218:Myosins / Q000235:genetics*; D009363:Neoplasm Proteins; D016133:Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics*; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D009363:Neoplasm Proteins; D014157:Transcription Factors; D009218:Myosins