Casticin is one of the main components from Fructus Viticis, which is widely used as an
anti-inflammatory agent. The mechanism of how
casticin affects
melanoma cell migration and invasion is still not well known. Here we studied the anti-
metastasis effects of
casticin on A375.S2
melanoma cells by using a non-lethal concentration. First; we used an adhesion assay to test the A375.S2 cells' adhesion ability
after treatment with
casticin. We next investigated the cell migration ability after
casticin treatment by using a wound healing assay to prove that the migration of A375.S2 cells can be inhibited by
casticin and double checked the results using the transwell-migration assay. The suppressive effects on
matrix metalloproteinase-2; and -9 (
MMP-2; and -9) activities were examined by
gelatin zymography. Furthermore, western blotting was used to investigate the
protein level changes in A375.S2 cells. We found that p-EGFR; Ras and p-ERK1/2 are decreased by
casticin, indicating that
casticin can down-regulate the migration and invasion ability of A375.S2 cells via the p-EGFR/Ras/p-ERK pathway. The NF-κB p65 and p-ERK levels in
nuclear proteins are also decreased by treatment with
casticin. An EMSA assay also discovered that the NF-κB p65 and
DNA interaction is decreased. NF-κB p65
protein level was examined by immunofluorescence staining and also decreased. Our findings suggest that
casticin has anti-metastatic potential by decreasing the invasiveness of A375.S2 cells. We also found that
casticin suppressed A375.S2 cell proliferation and cell adhesion ability, but did not affect cell death, as examined using cytometry and a
collagen adhesion assay. Based on these observations,
casticin could be used as an inhibitor of migration and invasion of human
melanoma cells in the future.