Detection of PAX8-PPARG fusion transcripts in archival thyroid carcinoma samples by conventional RT-PCR

Genes Chromosomes Cancer. 2012 Apr;51(4):402-8. doi: 10.1002/gcc.21925. Epub 2011 Dec 16.

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.

MeSH terms

  • DNA Primers
  • Genetic Variation
  • Humans
  • Nucleic Acid Amplification Techniques / methods
  • Oncogene Proteins, Fusion / analysis
  • Oncogene Proteins, Fusion / genetics*
  • PAX8 Transcription Factor
  • PPAR gamma / genetics*
  • Paired Box Transcription Factors / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Thyroid Neoplasms / genetics*

Substances

  • DNA Primers
  • Oncogene Proteins, Fusion
  • PAX8 Transcription Factor
  • PAX8 protein, human
  • PPAR gamma
  • Paired Box Transcription Factors