Gene expression profiling of potential peroxisome proliferator-activated receptor (PPAR) target genes in human hepatoblastoma cell lines inducibly expressing PPAR isoforms

Keisuke Tachibana, Yumi Kobayashi, Toshiya Tanaka, Masayuki Tagami, Akira Sugiyama, Tatsuya Katayama, Chihiro Ueda, Daisuke Yamasaki, Kenji Ishimoto, Mikako Sumitomo, Yasutoshi Uchiyama, Takahide Kohro, Juro Sakai, Takao Hamakubo, Tatsuhiko Kodama, Takefumi Doi

Research output: Contribution to journalArticlepeer-review

94 Citations (Scopus)

Abstract

Background: Peroxisome proliferator-activated receptors (PPARs) are ligand-activated-transcription factors and commonly play an important role in the regulation of lipid homeostasis. To identify human PPARs-responsive genes, we established tetracycline-regulated human hepatoblastoma cell lines that can be induced to express each human PPAR and investigated the gene expression profiles of these cells. Results: The expression of each introduced PPAR gene was investigated using the various concentrations of doxycycline in the culture media. We found that the expression of each PPAR subtype was tightly controlled by the concentration of doxycycline in these established cell lines. DNA microarray analyses using these cell lines were performed with or without adding each subtype ligand and provided much important information on the PPAR target genes involved in lipid metabolism, transport, storage and other activities. Interestingly, it was noted that while ligand-activated PPARδ induced target gene expression, unliganded PPARδ repressed these genes. The real-time RT-PCR was used to verify the altered expression of selected genes by PPARs and we found that these genes were induced to express in the same pattern as detected in the microarray analyses. Furthermore, we analysed the 5′-flanking region of the human adipose differentiation-related protein (adrp) gene that responded to all subtypes of PPARs. From the detailed analyses by reporter assays, the EMSAs, and ChIP assays, we determine the functional PPRE of the human adrp gene. Conclusion: The results suggest that these cell lines are important tools used to identify the human PPARs-responsive genes.

Original languageEnglish
Article number3
JournalNuclear Receptor
Volume3
DOIs
Publication statusPublished - 2005 Oct 3
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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