Figure 5. uPA mediates HDAI-induced invasion in human cancer cells.

A. The invasive capacity of SK-N-BE, SK-N-AS, SF-3061 and LNCaP cells treated with TSA, NaB or SCR was determined in vitro using the matrigel invasion assay. Cells invading through the matrigel were counted under a microscope in five random fields at a 200X magnification. Each bar represents the mean ± SD of three fields where significant differences between control cells, which exhibited undetectable uPA mRNA expression, and treated cells are represented by asterisks* (p<0.05).

B. LNCaP cells stably transfected with control or uPA shRNA were subjected to matrigel invasion assays for 24 h in the absence (mock) or presence of TSA, NaB or SCR (top). Asterisks (*) show significant differences between controls and treatment groups (p <0.05). The mRNA expression of uPA in LNCaP cells with the indicated treatment was detected by RT-PCR (bottom). Numbers indicated below each band are the quantitative values from densitometric analyses. Data are normalized to corresponding coamplified GAPDH level, averaged and represented as percent of no shRNA control (no shRNA set to one).

C. Immunoblot (top) and RT-PCR (bottom) analysis of LNCaP cells stably transfected with a vector expressing uPA, empty vector or mock. GAPDH was used as a loading control for RNA and protein analyses.

D. The invasive capacity of cells stably expressing uPA was examined in vitro by matrigel invasion assay. Significant differences of the results obtained for the transfectants and for the LNCaP control cells are indicated by asterisks* (p<0.05).

(All results are representative of three separate experiments.)