RNA interference against peroxisome proliferator-activated receptor delta gene promotes proliferation of human colorectal cancer cells

Dis Colon Rectum. 2008 Mar;51(3):318-26; discussion 326-8. doi: 10.1007/s10350-007-9145-8. Epub 2008 Jan 24.

Abstract

Purpose: This study was designed to investigate the effects of peroxisome proliferator-activated receptor delta (PPAR delta) on the proliferation and apoptosis of human colorectal cancer cells.

Methods: For RNA interfering (RNAi), HCT-116 cells were transfected with short hairpin RNA (shRNA)-expressing plasmids against PPAR delta or negative control vectors, and the stably transfected cells were selected with G418. The efficacy of RNAi was assessed by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting analysis. The proliferation, cell cycle, and apoptosis of HCT-116 cells treated by RNAi, compared with those containing control vectors or untreated, were analyzed respectively by using MTT (methyl thiazolyl tetrazolium), flow cytometry, and TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick end-labeling (TUNEL) assay.

Results: RNAi targeting PPAR delta resulted in substantial suppression of PPAR delta expression and significantly promoted the proliferation of HCT-116 cells relative to those with control vectors or untreated, obviously decreasing the frequency of G1-phase cells but had no effect on cell apoptosis.

Conclusions: PPAR delta may inhibit the proliferation of CRC cells and increase the number of cells in G1 phase, without any function in cell apoptosis.

MeSH terms

  • Analysis of Variance
  • Apoptosis
  • Blotting, Western
  • Cell Cycle / drug effects
  • Cell Line, Tumor
  • Cell Proliferation*
  • Colorectal Neoplasms / genetics
  • Colorectal Neoplasms / metabolism*
  • Flow Cytometry
  • Humans
  • In Situ Nick-End Labeling
  • PPAR delta / genetics*
  • Plasmids
  • RNA Interference*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction

Substances

  • PPAR delta