Detection of myc translocations in lymphoma cells by fluorescence in situ hybridization with yeast artificial chromosomes.
Translocations involving chromosome 8 at band q24 and one of the Ig loci on chromosomes 14q32, 22q11, and 2p11 are the hallmark of Burkitt's lymphoma (BL). It has been previously observed that the exact localization of the breakpoints at chromosome 8q24 can vary significantly from patient to patient, scattering over a distance of more than 300 kb upstream of c-myc and about 300 kb downstream of c-myc. To generate probes for fluorescence in situ hybridization (FISH) that detect most c-myc translocations, we screened a yeast artificial chromosome (YAC) library from normal human lymphocytes by colony hybridization, using three markers surrounding the c-myc gene as probes. We obtained 10 YAC clones ranging in size between 500 and 200 kb. Two nonchimeric clones were used for FISH on several BL cell lines and patient samples with different breakpoints at 8q24. Our results show that the YAC clones detected translocations scattered along approximately 200 kb in both metaphase chromosomes and interphase nuclei. The sensitivity, rapidity, and feasibility in nondividing cells render FISH an important diagnostic tool. Furthermore, the use of large DNA fragments such as YACs greatly simplifies the detection of translocations with widely scattered breakpoints such as these seen in BL.
Veronese
M L
ML
Jefferson Cancer Institute, Jefferson Medical College, Philadelphia, PA 19107, USA.
Ohta
M
M
Finan
J
J
Nowell
P C
PC
Croce
C M
CM
D000328:Adult; D002051:Burkitt Lymphoma / Q000235:genetics* / Q000473:pathology; D002648:Child; D002675:Child, Preschool; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D002883:Chromosomes, Human, Pair 14 / Q000648:ultrastructure*; D002892:Chromosomes, Human, Pair 22 / Q000648:ultrastructure*; D002898:Chromosomes, Human, Pair 8 / Q000648:ultrastructure*; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D005803:Genes, Immunoglobulin; D016259:Genes, myc; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016400:Lymphoma, Large-Cell, Immunoblastic / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D012680:Sensitivity and Specificity; D014178:Translocation, Genetic; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm