Involvement of Fatty Acid Binding Protein 5 and PPAR/ in Prostate Cancer Cell Growth
Figure 3
FABP5 is a direct target gene for PPAR. (a) PC3M cells were treated with cycloheximde (20g/mL) for 10 min prior to addition of GW0742 (1 M, 4 h). Expression of FABP5 mRNA was measured by Q-PCR. Data are meanS.D. (). * versus nontreated control. (b) Location of putative PPREs in the FABP5 promoter. (c) Transcriptional activation assays utilizing a luciferase reporter driven by a 900 bp region of the FABP5 promoter which encompasses the four putative PPREs (5 promoter). In the mutant reporter construct (5 promoter/PPRE4m), the AGCTCA sequence in PPRE4 was exchanged to AGCTTT. PC3M cells were cotransfected with the denoted reporter and an expression vector for -galactosidase. 24 h later, cells were treated with vehicle or GW0742 and cultured for 24 hr. Luciferase activity was measured and normalized to the activitiy of b-galactosidase. Data are meanS.D. (). Inset: Luciferase assays carried out with a luciferase reporter driven by 3 consensus PPREs. (d) Chromatin immunoprecipitation assays in denoted cell lines. Immunoprecipitations were carried out using denoted antibodies and the PPRE4 region of the FABP5 promoter amplified by PCR. Bottom panel: quantification of 3 independent ChiP assays (meanSEM).
