A break-apart fluorescence in situ hybridization assay for detecting RET translocations in papillary thyroid carcinoma.
At least 15 different translocations have been described activating RET in papillary thyroid carcinomas. A break-apart fluorescence in situ hybridization (FISH) assay should detect many translocations involving the RET gene without requiring knowledge of the partner gene. G-banding and spectral karyotyping was performed to further characterize the papillary carcinoma cell line TPC-1. BAC clones flanking the 5' and 3' regions of RET were labeled with SpectrumRed and biotin detected with avidin-AMCA (blue). In addition to the previously described chromosomal t(1;10;21), TPC-1 was found to have del(7)(q22q31) and der(8)t(8;8)(p21;q11.2). With the BAC probes, TPC-1 interphase nuclei showed the expected signal pattern of one paired red-blue signal as well as separated red and blue signals from the rearranged RET gene in 93% of cells. Interphase nuclei from normal human lymphocytes showed two paired red-blue signals in 97% of cells. TPC-1 cells were found to have the previously described chromosomal rearrangement that involves chromosome 10, with few other cytogenetically detectable genomic alterations. RET rearrangement can be detected by a break-apart BAC FISH probe set in the interphase nuclei of TPC-1 cells. This assay can potentially detect clinically relevant translocations involving RET.
Chen
Frank
F
Department of Pathology, The Johns Hopkins Hospital, Baltimore, MD 21287, USA.
Clark
Douglas P
DP
Hawkins
Anita L
AL
Morsberger
Laura A
LA
Griffin
Constance A
CA
D002291:Carcinoma, Papillary / Q000235:genetics*; D045744:Cell Line, Tumor; D002871:Chromosome Banding; D002874:Chromosome Mapping; D002879:Chromosomes, Human, Pair 10; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013964:Thyroid Neoplasms / Q000235:genetics*; D014178:Translocation, Genetic / Q000235:genetics*
D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human