Figure 3

Betulin and its derivatives inhibit cell migration in the MDA-MB-231 cell line. A wound healing assay was performed to determine the cell migration ability. MDA-MB-231 cells were treated with 0.1% DMSO (A, B, C) or 5 μg/mL K02 (D, E, F), K03 (G, H, I), K04 (J, K, L), or K06 (M, N, O) for 24 h. The cells were observed every 12 h under the same observation field for wound healing status. At 12 h, cell migratory abilities in K03- (H) and K04-treated (K) cells slightly decreased compared with DMSO controls (B). In K02- (F) and K06-treated (N) cells, cell migratory abilities more obviously decreased compared with DMSO controls (B). At 24 h, cell migratory abilities in the K02 (L), K03 (H), K04 (K), and K06 (O) groups significantly decreased compared with DMSO controls (B), and the wound area was larger than that of DMSO controls (C). The wound area was quantified by ImageJ software. Compared with 12 h of DMSO treatment, the wound area was slightly significantly larger (* p < 0.05) in the K03- and K04-treated groups and more significantly larger in the K02- and K06-treated groups (** p < 0.01). Compared with 24 h of DMSO treatment, the wound area was significantly larger (# p < 0.05) in the K03- and K04-treated groups and also significantly larger in the K02- and K06-treated groups (## p < 0.01).