title: Detection of PAX8-PPARG fusion transcripts in archival thyroid carcinoma samples by conventional RT-PCR.
abstract: The t(2;3)(q13;p25) occurs in a subgroup of follicular-patterned thyroid tumors and leads to a fusion of the genes encoding for the thyroid-specific transcription factor paired box 8 (PAX8) and the peroxisome proliferator-activated receptor gamma (PPARγ). Although initially discovered in follicular carcinomas (FTC), the fusion transcripts were also detected in a small fraction of follicular adenomas and rarely in follicular variants of papillary carcinomas (FV-PTC). In most RT-PCR based studies, fresh or snap-frozen tissue samples were used. The aim of the present study was to develop a method for the detection of chimeric PAX8-PPARG transcripts in formalin-fixed paraffin-embedded (FFPE) thyroid tumor samples by conventional RT-PCR. For this purpose, RNA from FFPE samples of 21 FTC, seven FV-PTC, and one bone metastasis derived from an FTC was subjected to RT-PCR with subsequent gel electrophoretic separation of the products. Fusion transcripts were detected in 2/21 primary FTC (9.5%) and in the bone metastasis, but they were undetectable in all seven FV-PTC under investigation. The RT-PCR approach described herein allows to detect all known variants of PAX8-PPARG fusion transcripts and is applicable to FFPE tissues. Thus, it can be used to screen archival thyroid tumor samples for the gene fusion.
authors.affiliation: Center for Human Genetics, University of Bremen, Bremen, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Klemke
authors.forename: Norbert
authors.initials: N
authors.lastname: Drieschner
authors.forename: Gazanfer
authors.initials: G
authors.lastname: Belge
authors.forename: Käte
authors.initials: K
authors.lastname: Burchardt
authors.forename: Klaus
authors.initials: K
authors.lastname: Junker
authors.forename: Jörn
authors.initials: J
authors.lastname: Bullerdiek
mesh_terms: D017931:DNA Primers; D014644:Genetic Variation; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D015514:Oncogene Proteins, Fusion / Q000032:analysis / Q000235:genetics*; D000072636:PAX8 Transcription Factor; D047495:PPAR gamma / Q000235:genetics*; D051761:Paired Box Transcription Factors / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013964:Thyroid Neoplasms / Q000235:genetics*
chemical_list: D017931:DNA Primers; D015514:Oncogene Proteins, Fusion; D000072636:PAX8 Transcription Factor; C083325:PAX8 protein, human; D047495:PPAR gamma; D051761:Paired Box Transcription Factors