A long distance-PCR derived FISH probe detects a deletion between p15 and p16 in CML and T-ALL patients.
The tumor suppressor genes p15INK4B and p16INK4A, located in the chromosomal region 9p21, are frequently inactivated by homo- or hemizygous deletions, point mutation or promotor methylation in various types of cancer. No commercial probe is yet available that allows the detection of such deletions by FISH. Long distance (LD)-PCR was successfully used to generate a FISH probe, that covers a sequence stretch of 11.68 kb, located between the tumor suppressor genes p15 and p16. The LD-PCR amplicon was cloned and biotinylated by DOP-PCR (degenerated oligonucleotide primed-PCR) or nick translation. The FISH probe was hybridized on different samples of 16 patients with leukemia (3 T-ALL, 13 CML) and normal controls. Loss of at least one FISH-signal was found in 2/3 (67%) of the T-ALL- and 2/13 (15%) of the CML-cases. The new FISH probe presented here was proven to be advantageous for the detection of deletions in chromosomal region 9p21, especially between p15 and p16.
Bleichert
A
A
Institute of Human Genetics and Anthropology, Jena, Germany.
Fiedler
W
W
Claussen
U
U
Ernst
G
G
Loncarevic
I F
IF
Heller
A
A
Liehr
T
T
Kunert
C
C
von Eggeling
F
F
D000293:Adolescent; D000328:Adult; D000368:Aged; D001752:Blast Crisis / Q000235:genetics; D016022:Case-Control Studies; D002648:Child; D002872:Chromosome Deletion; D002899:Chromosomes, Human, Pair 9; D019175:DNA Methylation; D005260:Female; D006720:Homozygote; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008957:Models, Genetic; D015335:Molecular Probes; D017354:Point Mutation; D016133:Polymerase Chain Reaction
D015335:Molecular Probes