Fig. 3
DICER1 silencing increases proliferation, migration and invasion in vitro. a–f Cal62 and TPC1 cell lines were transfected with an siRNA against DICER1 (siDICER1) or a control siRNA (siControl) and analysis was performed 48 h later. a Relative expression levels of miRNAs. b Immunoblot of DICER1 and proliferating cell nuclear antigen (PCNA). Actin was used as a loading control. c Representative images of crystal violet-stained cells. d BrdU incorporation relative to siControl-transfected cells. e Representative images from a wound healing assay at 0 and 16 h (Cal62, left) or 0 and 48 h (TPC1, right) after scratching. f Quantification of the invasion rates. Left: representative images of the lower chamber (invading cells). Right: cell invasion rates relative to siControl cells. g, h Cells were transfected with an siRNA against DICER1 (siDICER1) or a control siRNA (siControl). g Relative mRNA levels assayed by qRT-PCR of EMT genes CDH1, EYA1, EYA2, FN, PAI1, SNAIL1, TWIST1, and ZEB1 72 h after siRNA transfection with RNAiMAX Lipofectamine. h Immunoblot for DICER1 and TWIST1 (left) and DICER1, ZEB1, and fibronectin (right) 48 h after siRNA transfection. Actin was used as a loading control. Values represent mean ± SD (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001
