Detection of 9p deletions in leukemia cell lines by interphase fluorescence in situ hybridization with YAC-derived probes.
Hemizygous and homozygous deletions of the type I interferon gene cluster (IFN) have been detected in about 20% of acute lymphoblastic leukemias. A putative tumor suppressor gene (TSG) is thought to be located centromeric to the IFN cluster on chromosomal bands 9p21-22. We studied the accuracy of fluorescence in situ hybridization (FISH) for detecting deletions in interphase cells using yeast artificial chromosome (YAC) clones containing all or part of the IFN cluster. FISH probes were generated from YACs (320-1300 kb in size) by a sequence-independent amplification technique (SIA). Fifteen cell lines (nine T-ALL, three B-cell precursor ALL, one B-ALL, one AML, one CML-BC) that had been well characterized by conventional cytogenetic analysis and molecular techniques were analyzed. We were able to detect all numerical changes of the IFN cluster including homozygous and hemizygous deletions accurately and to define subclones of the cell lines. Moreover, in six cell lines we were able to identify subclones. In dilution experiments the detection thresholds for subpopulations with homozygous and hemizygous deletions were determined to be 5% and 7.5%, respectively.
Dreyling
M H
MH
Department of Medicine, University of Chicago, Illinois 60637, USA.
Kobayashi
H
H
Olopade
O I
OI
Le Beau
M M
MM
Rowley
J D
JD
Bohlander
S K
SK
D001483:Base Sequence; D002872:Chromosome Deletion; D018244:Chromosomes, Artificial, Yeast; D015342:DNA Probes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007938:Leukemia / Q000235:genetics*; D008677:Metaphase; D008969:Molecular Sequence Data; D011003:Ploidies; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D015342:DNA Probes