Figure 4.

CHB inhibits wound closure, invasion, and migration in B16F0 cells. B16F0 cells were treated with CHB (0.5, 1.0, and 1.5 μg/mL). (A) The inhibition rate of B16F0 cells was determined by SRB assay. (B) Adhesion ability test. (C, magnification, x200) Typical pictures of CHB on migration ability. (D) The analysis of the migrated B16F0 cells by quantitation. (E,F, magnification, x200), Migration and invasion of B16F0 cells were detected using the transwell assay. Relative invasion ability was determined by counting the cells on the lower surface of the filters in five individual fields. (G) Mmp2, Mmp9, Timp1, and Timp2 mRNA levels were analyzed by qPCR. Mouse Gapdh mRNA was used as the internal control. All mRNA levels were normalized to the mean value of the control group. (H) Mmp2 and Mmp9 expressions were measured by western blots. (I) Band density of the specific protein was analyzed with Quantity One image software, and the results are expressed as average density to β-actin. Results were expressed as mean ± SD for three separate experiments. *P < 0.05, **P < 0.01 compared with control group cells.