Fig. 2

Panel a 1,25-dihydroxyvitamin D3 inhibits migration of human and murine germline-derived cell lines. Transmigration of a human ovarian cancer cell line (A2780), a human teratocarcinoma cell line (NTERA-2), and a murine embryonal teratocarcinoma cell line (P19) through Transwell membranes (8-μm pore size) in response to 1,25-dihydroxyvitamin D3 at the indicated concentrations. Cells were rendered quiescent in 0.5 % BSA in culture medium overnight at 37 °C. The effects of 1,25-dihydroxyvitamin D3 on migration of all cell lines employed (6 × 10⁴ cells/100 μL/insert) were also evaluated in parallel for migration in response to 10 % FBS and 0.5 % BSA plus vehicle as a positive and negative control, respectively. Twenty-four hours later, the migrated cells were stained and counted using an inverted microscope. Panel b 1,25-dihydroxyvitamin D3 interferes with the adhesiveness of human and murine germline-derived cell lines to fibronectin. Adhesion of the A2780 human ovarian cancer cell line, the NTERA-2 human teratocarcinoma cell line, and the P19 murine embryonal teratocarcinoma cell line to fibronectin-coated surfaces in response to 1,25-dihydroxyvitamin D3. After three hours of quiescence, cells (3000 cells/100 μL) were stimulated with 1,25-dihydroxyvitamin D3 at the indicated concentrations in medium with 0.5 % BSA for 5 min at 37 °C. After the non-adherent cells were removed by three consecutive washes with PBS, the number of adherent cells was measured by microscopic analysis. The effects of 1,25-dihydroxyvitamin D3 on adhesion of all cell lines employed were also evaluated for adhesion compared with stromal-derived factor 1 (300 ng/mL) and culture medium containing 0.5 % BSA plus vehicle as a positive and negative control, respectively. The negative control values are normalized to 100 %. Data are displayed as means ± SD, with a statistical significance *p ≤ 0.05 versus control (unstimulated) cells