Figure 5. PIT-1 inhibits cancer cell migration and invasion.

(A) PIT-1 inhibits cancer cell migration in a transwell assay. SUM159 cells were treated with PIT-1 for 8 hr. The cells on the lower side of chamber were stained and the representative images are shown, then cells were lysed and colorimetric determination was made at 595 nm.

(B) PIT-1 inhibits cancer cell migration in a wound healing assay. A scratch was introduced into a monolayer of SUM159 cells, followed by treatment with PIT-1 for 8 hr. The width of wounded cell monolayer was measured in five random fields, and the representative images are shown.

(C) Quantitation of the data from the assays in (A) and (B).

(D,E) PIT-1 and PIT-2, but not PIT-1i-1 and PIT-1i-2, inhibit SUM159 cell migration in transwell (D) and wound healing (E) assays.

(F) PIT-1 inhibits cancer cell invasion. SUM159 cells were seeded on a matrigel pre-coated transwell membrane, and the treatment and analysis are similar with transwell assay described above.

(G) PIT-1 and PIT-2, but not PIT-1i-1 and PIT-1i-2, inhibit cancer cell invasion through a matrigel-coated membrane.

(H) PIT-1 reversibly inhibits acquisition of the invasive phenotype of SUM159 cells in a 3-D matrigel matrix. SUM159 cells were seeded in matrigel and incubated for 192 hr, followed by the treatment with PIT-1 (12.5 μM) for 114 hr. Subsequently, PIT-1 was washed out and cells were incubated for additional 96 hr. The representative images are shown.