Figure 8

EpCAM expression modulates expression of AP-1 target genes known to be involved in cancer invasion. MDA-231 cells were transiently transfected with control and epithelial cell adhesion molecule (EpCAM) cDNA expression constructs and allowed to stabilize for 24 hours. The cells were then serum-starved for 16 hours, and then stimulated with 10% FBS DMEM growth media for two hours. RNA was extracted using RNeasy (Qiagen). Two micrograms of RNA was reverse transcribed and 40 ng equivalent cDNA was used for each assay in triplicate. Activator protein 1 (AP-1) target genes CTSL1, Autotaxin, and ARP2/3 were analyzed using real-time PCR. Relative expression was quantified using the [delta] [delta] Ct method with 18S rRNA as a control. The results are representative of at least two independent experiments.