The Function and Significance of SELENBP1 Downregulation in Human Bronchial Epithelial Carcinogenic Process
Figure 3
The effects of SELENBP1 gene knockout on the B[a]P-induced human bronchial epithelial cell transformation.
A, Western blotting shows SELENBP1 expression in the untransfected (1), empty vector pLKO.1-transfected (2, 3) and pLKO.1-SELENBP1-shRNA-tansfected 16HBE cells (4, 5). β-actin is used as an internal control for loading. B, cell growth in low serum medium after exposed to B[a]P for 18 weeks. Cells were subjected to MTT assay as described in “Materials and Methods”. Three experiments were done; points, mean; bars, S.D. (*, P<0.05 vs. untransfected or empty vector -transfected 16HBE cells after exposed to B[a]P by Student’s t test). C, anchorage dependent colony growth after cells exposed to B[a]P for 18 weeks. (left) cells were subjected to plate colony formation assay as described in “Materials and Methods”, and colonies were stained with crystal violet and photographed under camera; (right) the histogram showed plate colony formation rates. Three experiments were done; columns, mean; bars, S.D. (**, P<0.05 vs. untransfected or empty vector -transfected 16HBE cells after exposed to B[a]P by One-way ANOVA). D, anchorage independent colony growth after cells exposed to B[a]P for 18 weeks. (left) cells were subjected to soft agar colony formation assay as described in “Materials and Methods”, and colonies were photographed under microscope; (right) the histogram showed number of soft agar colonies in 10 randomly chosen microscopic fields using a 5× objective. Three experiments were done; columns, mean; bars, S.D. (*, P<0.05 vs. untransfected or empty vector -transfected 16HBE cells after exposed to B[a]P by One-way ANOVA). Cell proliferation, and plate and soft agar colony growth of the cells exposed to vehicle (DMSO) for 18 weeks are also shown and used as controls. 16HBE, untransfected cells; 16HBE/pLKO.1, empty vector pLKO.1-transfected cells; 16HBE/pLKO.1-SELENBP1-shRNA, pLKO.1-SELENBP1-shRNA -tansfected cells.