Antidiabetic thiazolidinediones induce ductal differentiation but not apoptosis in pancreatic cancer cells

Figure 2 PPARγ expression and transcriptional activity in HPAC cells transduced with PPARγ-expressing retrovirus. A: HPAC cells were stable transduced by retrovirus driving expression of human PPARγ (hPPARγ-pLNCX) as described in methods. After selection with G418 four resistant clones were expanded and screened for hPPARγ expression. The cell extracts were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted to nitrocellulose. The proteins (40 μg) were detected with antibody raised against human PPARγ. P1-P4 represent nuclear protein extracted from G418-resistant HPAC cell clones; M represents nuclear proteins from mock (pLNCX) transduced HPAC cells; W (wild type) represents nuclear proteins from untransduced, parental HPAC cells; B: After overnight attachment cells were transfected with ARE-7₃-tk-luciferase reporter plasmid and pSV₂-CAT as internal control for transfection efficiency. Twenty-four hours after transfection cells were treated with RGZ at the indicated concentration. Twenty-four hours after treatment the cells were harvested for luciferase and CAT assay as described in Methods. The data is expressed as mean±SD for 4 replicate experiments performed in triplicate; ^aP<0.05 vs control; C: Effect of TDZ treatment on anchorage-independent growth of HPAC cells transduced with PPARγ-expressing retrovirus. Clonogenic assay of P3-HPAC cells treated with the indicated concentration of RGZ was performed as described in materials and methods. The number of colonies was then given as the percentage of control cells treated with vehicle alone. The mean±SD of five independent experiments performed for each in triplicate are shown. ^aP<0.05 vs Mock transduced cells.