Figure 7.

Roles of podocalyxin‐like protein (PODXL) and gelsolin in the motility and invasiveness of pancreatic ductal adenocarcinoma cells. (a,b) Oligonucleotides (siRNAs targeting gelsolin [siGelsolin] or scrambled RNAs [Scr] as the negative control) were transiently transfected into S2‐013 and PANC‐1 cells. Motility (a) and two‐chamber invasion assays (b) were carried out. Migrating cells in four fields per group were scored. Data are derived from three independent experiments. Columns, mean; bars, SD. *P < 0.004 and **P < 0.002 compared with Scr‐transfected S2‐013 cells (Student's t‐test). (c) Expression of endogenous PODXL and gelsolin in PDAC cells (PANC‐1, S2‐013, and BxPc‐3) compared with the HPNE cell line, as determined by Western blotting. (d,e) A myc‐tagged PODXL‐rescue construct was transfected into S2‐013 (d) and PANC‐1 (e) cells that had been transfected with PODXL siRNA and gelsolin siRNA; 48 h later, motility and two‐chamber invasion assays were undertaken. Migrating cells in four fields per group were counted. Data are derived from three independent experiments. Columns, mean; bars, SD. *P < 0.001 compared with corresponding PODXL siRNA and gelsolin siRNA‐transfected S2‐013 cells that were transfected with mock vector (Student's t‐test). (f) Mock control vector or myc‐tagged PODXL‐rescue construct were transiently transfected into BxPc‐3 (left panel) and PANC‐1 (right panel) cells that had been transfected with PODXL‐siRNA; 48 h later, motility and two‐chamber invasion assays were carried out. Migrating cells in four fields per group were counted. Data are derived from three independent experiments. Columns, mean; bars, SD. *P < 0.006 compared with PODXL siRNA‐transfected PANC‐1 cells that were transfected with mock vector (Student's t‐test).