Rhinacanthin-C Extracted from Rhinacanthus nasutus (L.) Inhibits Cholangiocarcinoma Cell Migration and Invasion by Decreasing MMP-2, uPA, FAK and MAPK Pathways

摘要

Cholangiocarcinoma is a malignant tumor with high metastatic and mortality rates. We investigated the effectsof rhinacanthin-C on cell proliferation, migration, invasion and the expression of proteins regulating cancer cellinvasion-regulated proteins in a cholangiocarcinoma (KKU-M156) cell line. Cytotoxicity of rhinacanthin-C wasdetermined by the SRB assay. Using wound-migration, chamber-migration and chamber-invasion assays, we assessedthe effects of rhinacanthin-C against KKU-M156 cells. The activities of matrix metalloproteinases 2 and 9 (MMP-2,MMP-9) and urokinase-type plasminogen activator (uPA) were determined using gelatinase and uPA zymographyassays. The expression of invasion-regulated proteins was investigated using western-blot analysis. After treatmentwith rhinacanthin-C, KKU-M156 cells exhibited antiproliferative effects in a dose-dependent manner with greaterefficacy than in Vero cells: IC50 values were 1.50 and 2.37 μM, respectively. Rhinacanthin-C significantly inhibited cellmigration and invasion of KKU-M156 cells in a dose-dependent manner. Consistent with this observation, treatmentwith rhinacanthin-C was associated with a decrease in the expression levels of FAK, p-FAK, MMP-2, and a decrease inthe levels of p38-, JNK1/2- and ERK1/2-MAPK pathways as well as inhibiting NF-κB/p65 expression and translocationof NF-κB/p65 to the nucleus. We have shown for the first time that the anti-metastatic effects of rhinacanthin-C onKKU-M156 cells are mediated via inhibition of the expression of invasion-regulated proteins. Rhinacanthin-C maydeserve consideration as a potential agent for the treatment of cholangiocarcinoma.