HAX1 Augments Cell Proliferation, Migration, Adhesion, and Invasion Induced by Urokinase-Type Plasminogen Activator Receptor
Figure 3
HAX1 overexpression increases cell migration and adhesion in uPAR-stimulated cells. (a) HEK293/uPAR, (b) HCT116, and (c) MDA-MB-231 cells were transfected with pGEM-3Zf(+) (light bars) or pGEM-3Zf(+)∖HAX1 (dark bars) and examined using wound healing assay. A monolayer of confluent cells was scratched with a 200 μL pipette tip and the closure of the scratch was observed in the presence of either 10% FCS, 1% FCS, or 100 ng/mL of EGF, uPA, or ATF. Photographs were taken immediately and after 18 h of creating the scratch. Cell migration was assessed by measuring the distance migrated by cells in pixels in these 18 h. (d) HAX1 augments HEK293/uPAR cell adhesion. 1.5 × 105 cells/well transfected with pGEM-3Zf(+) (light bars) or pGEM-3Zf(+)∖HAX1 (dark bars) were seeded in precoated 96-well plate with 2 μg/mL Vn or heat-denatured BSA, treated with 100 ng/mL of EGF, uPA, or uPA-ATF, and incubated for 2 h at 37°C. After washing, the attached cells were fixed with methanol/acetone and stained with 0.1% crystal violet. The stain was eluted using acetic acid/methanol/water and absorbance measured at O.D595. Bars are mean ± S.E using unpaired Student’s t-test (n.s; *P < 0.05; **P < 0.01; ***P < 0.001).
