Genomic binding and regulation of gene expression by the thyroid carcinoma-associated PAX8-PPARG fusion protein

Oncotarget. 2015 Dec 1;6(38):40418-32. doi: 10.18632/oncotarget.6340.

Abstract

A chromosomal translocation results in production of an oncogenic PAX8-PPARG fusion protein (PPFP) in thyroid carcinomas. PAX8 is a thyroid transcription factor, and PPARG is a transcription factor that plays important roles in adipocytes and macrophages. PPFP retains the DNA binding domains of both proteins; however, the genomic binding sites of PPFP have not been identified, and only limited data exist to characterize gene expression in PPFP thyroid carcinomas. Therefore, the oncogenic function of PPFP is poorly understood. We expressed PPFP in PCCL3 rat thyroid cells and used ChIP-seq to identify PPFP genomic binding sites (PPFP peaks) and RNA-seq to characterize PPFP-dependent gene expression. PPFP peaks (~20,000) include known PAX8 and PPARG binding sites and are enriched with both motifs, indicating that both DNA binding domains are functional. PPFP binds to and regulates many genes involved in cancer-related processes. In PCCL3 thyroid cells, PPFP binds to adipocyte PPARG target genes in preference to macrophage PPARG target genes, consistent with the pro-adipogenic nature of PPFP and its ligand pioglitazone in thyroid cells. PPFP induces oxidative stress in thyroid cells, and pioglitazone increases susceptibility to further oxidative stress. Our data highlight the complexity of PPFP as a transcription factor and the numerous ways that it regulates thyroid oncogenesis.

Keywords: differentiation; follicular thyroid cancer; gene fusion; peroxisome proliferator-activated receptor gamma; pioglitazone.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Biomarkers, Tumor / genetics*
  • Chromatin Immunoprecipitation
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic*
  • Genome / genetics*
  • High-Throughput Nucleotide Sequencing / methods
  • Hypoglycemic Agents / pharmacology
  • Oncogene Proteins, Fusion / genetics*
  • PAX8 Transcription Factor
  • PPAR gamma / genetics*
  • Paired Box Transcription Factors / genetics*
  • Pioglitazone
  • RNA, Messenger / genetics
  • Rats
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thiazolidinediones / pharmacology
  • Thyroid Neoplasms / drug therapy
  • Thyroid Neoplasms / genetics*
  • Thyroid Neoplasms / metabolism
  • Tumor Cells, Cultured

Substances

  • Biomarkers, Tumor
  • Hypoglycemic Agents
  • Oncogene Proteins, Fusion
  • PAX8 Transcription Factor
  • PAX8 protein, human
  • PPAR gamma
  • Paired Box Transcription Factors
  • RNA, Messenger
  • Thiazolidinediones
  • Pioglitazone