Anti-proliferation, pro-apoptosis and anti-migration

Ginkgolic acids (GA) have been reported to exhibit anticancer properties, however, 15 the mechanisms remain unclear. This study aims to investigate the mechanisms of GA C13:0 16 that was isolated from Ginkgo biloba exocarp (GBE) for anti-proliferation, pro-apoptosis and 17 anti-migration effects in human MCF-7 and mouse 4T-1 breast cancer cells. The cytotoxic 18 effect, apoptosis induction and migration inhibition were measured using MTT, TUNEL and 19 Wound healing assays. The expression of mRNA and protein were determined using qPCR 20 and Western blot. Our results showed that no cytotoxicity was found at concentrations of 21 C13:0 below 100μM. The effects of GA C13:0 was further demonstrated by up-regulation of 22 the Bax/Bcl-2 apoptosis pathway and the expression of Apaf-1 protein in the mitochondria. In 23 addition, GA C13:0 also suppressed cell migration and epithelial to mesenchymal transition 24 (EMT) with the increase of E-cadherin expression accompanied by the decrease of Snail, 25 MMP-2, MMP-9 and Vimentin expression. Moreover, GA C13:0 induced cytochrome P450 26 (CYP) 1B1 expression in aryl hydrocarbon receptor (AhR) pathway. Notably, the 27 up-regulation of CYP1B1 also might play a pivotal regulatory role in mitochondrial and EMT 28 pathways in MCF-7 and 4T-1 cells. Our results may have implications for the development of 29 anticancer agents containing GA as functional additives. 30