Fig. 3
A p190A impairs invasion and migration capacities of BC cells both in in vitro and ex vivo experiments. (A) Relative mRNA and protein levels of p190A in normal ureter UR4-2, immortalized urothelial cell lines Y235T, HBLAK, low invasive bladder cancer cell lines RT4, BC61, and high invasive bladder cancer cell lines T24, UMUC3, J82, and BFTC. (B, C, D) Relative mRNA and protein levels of p190A in RT4-p190A-knockdown cells and BFTC/T24-p190A-overexpression cells. β-actin was used as a loading control. Representative images (E, G, I) and quantification (F, H, J) show that cell invasion is increased in RT4 cells with p190A knockdown and decreased in T24 and BFTC cells with p190A overexpression in Boyden chamber assay. Scale bar, 200 μm. Representative images (K, M, O) and quantification (L, N, P) show that cell migration is increased in RT4 cells with p190A knockdown and decreased in T24 and BFTC cells with p190A overexpression in would healing assay. Scale bar, 200 μm. Ex vivo porcine urinary bladder (PUB) model show that cell invasion is (Q) increased in RT4 cells with p190A knockdown and (R) decreased in T24 cells with p190A overexpression. The white arrow indicates cell invasion. The invasiveness of cells was detected by immunohistochemistry (IHC) staining with human leukocyte antigen (HLA) antibody. Scale bar, 100 μm and 50 μm. All experiments were performed at least in triplicates (n = 3). Statistical data are presented as mean ± SEM. ANOVA (for B, F, L) and Student’s t-test (for C, D, H, J, N, P) were applied for statistical analyses. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
