Figure 3.

Combination of TMZ and resveratrol (RES) prevented cell migration and inhibited cell metastasis. Representative pictures of SHG44 cells were at 0 h and 48 h after treatment (when a wound was made), and 24 h later, respectively. SHG44 cells subjected to in vitro scratch assay. The migration ability was estimated quantitatively by measuring the remaining area between the two edges of the wound. (A) At the beginning of 24 h, the gap distance was reduced to 1 μM in control groups and RES‐treated groups, and the gap distance of the TMZ group and the combined group was 83.2 ± 12.8 μM and 84.4 ± 26.4 μM, respectively (P= 0.8676). (B) During the last 24 h, the gap distance was reduced to 1 μM under control conditions, and after the treatment of RES and TMZ alone, the gap distance was reduced to 41.7 ± 8.8 and 87.3 ± 21.3 μM, indicating the partial inhibition of cell migration, and the most inhibitory effect after combination treatment was 304.1 ± 31.7 μM. The covered distance was estimated in five different areas of the culture, in three independent experiments. (C) Cells were incubated with indicated the treatment of TMZ at 100 μM and RES at 10 μM for 72 h, and then, cell proteins were obtained and analyzed with anti‐MMP‐9 antibodies by Western blot. The combination treatment significantly down regulated the expression of MMP‐9. *P < 0.05 versus control group (CON); ^#P < 0.05 versus TMZ.