Role of Peroxisome Proliferator-Activated Receptor and B-Cell Lymphoma-6 in Regulation of Genes Involved in Metastasis and Migration in Pancreatic Cancer Cells
Figure 2
Effect of activation on MMP-9 expression. (a) Miapaca-2 cells were transiently infected with nontargeting control, hBCL-6, , or hMMP9 lentiviral shRNAs for 48 hours. Total mRNA was isolated and gene-specific knockdown was assessed using qRT-PCR. *. (b) Miapaca-2 cells contain functional PPARs. Miapaca-2 cells were transiently infected with the indicated shRNAs and treated with the indicated PPAR isoform-specific agonists. Induction of the PPAR-target gene ADRP was determined using qRT-PCR. *. (c) Miapaca-2 cells were stimulated with human with or without GW501516 for 24 hours following transient infection with the indicated shRNAs. Human MMP-9 protein expression was quantified using MMP-9-specific ELISA (Invitrogen). *. (d) Miapaca-2 cells with reduced MMP-9 expression are less invasive than control Miapaca-2 cells. Following infection with human MMP-9-targeting shRNA, Miapaca-2 cells were seeded in 96-well invasion plates (Cell Biolabs, Inc.) and allowed to invade the basement membrane overnight. Relative cell invasion was quantified using the CytoSelect 96-well cell invasion assay with fluorometric readings at 480 nm/520 nm. *.
