title: Phytoestrogen Bakuchiol Exhibits In Vitro and In Vivo Anti-breast Cancer Effects by Inducing S Phase Arrest and Apoptosis.
abstract: Phytoestrogen has been proposed as an alternative to hormone replacement therapy, which has been demonstrated to promote a high risk of breast cancer. However, the effect of phytoestrogen on breast cancer development has not been fully understood. Bakuchiol is an active ingredient of a traditional Chinese herbal medicine Fructus Psoraleae, the dried ripe fruit of Psoralea corylifolia L. (Fabaceae). The in vitro and in vivo estrogenic activities and anti-breast cancer effects of bakuchiol have not been well-studied. We found that bakuchiol induced the GFP expression in transgenic medaka (Oryzias melastigma, Tg, Chg:GFP) dose-dependently (0-1 μg/ml), demonstrating its in vivo estrogenic activity. Low dose of bakuchiol (1 μg/ml) induced the cell proliferation and ERα expression in MCF-7 cells, which could be blocked by the anti-estrogen ICI 182780, suggesting the in vitro estrogenic activity of bakuchiol. Our data indicated that high doses of bakuchiol (>2 μg/ml) inhibited breast cancer cell growth, with a stronger anti-proliferative effect than resveratrol, a widely studied analog of bakuchiol. High doses of bakuchiol (4, 7, and 10 μg/ml) were used for the further in vitro anti-breast cancer studies. Bakuchiol induced ERβ expression and suppressed ERα expression in MCF-7 cells. It also induced S phase arrest in both MCF-7 and MDA-MB-231 cells, which could be rescued by caffeine. Knock-down of p21 also marginally rescued S phase arrest in MCF-7 cells. The S phase arrest was accompanied by the upregulation of ATM, P-Cdc2 (Tyr15), Myt1, P-Wee1 (Ser642), p21 and Cyclin B1, suggesting that blocking of Cdc2 activation may play an important role in bakuchiol-induced S phase arrest. Furthermore, bakuchiol induced cell apoptosis and disturbed mitochondrial membrane potential in MCF-7 cells. The bakuchiol-induced apoptosis was associated with increased expression of Caspase family and Bcl-2 family proteins, suggesting that bakuchiol may induce apoptosis via intrinsic apoptotic pathway. The in vivo anti-breast cancer effect of bakuchiol was further proved in zebrafish (Danio rerio, wild-type AB) xenografts. 0.5 μg/ml of bakuchiol significantly reduced the MCF-7 cell mass in zebrafish xenografts. Overall, these results suggested the potential of using bakuchiol in HRT and breast cancer treatment.
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong Hong Kong, China.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.affiliation: Vitargent (International) Biotechnology Limited Hong Kong, China.
authors.forename: Xueping
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong Hong Kong, China.
authors.forename: Chi C
authors.initials: CC
authors.lastname: Liu
authors.affiliation: Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University Hong Kong, China.
authors.forename: Lai S
authors.initials: LS
authors.lastname: Lee
authors.affiliation: Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University Hong Kong, China.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Man
authors.affiliation: Department of Biomedical Sciences, City University of Hong Kong Hong Kong, China.
authors.forename: Shuk H
authors.initials: SH
authors.lastname: Cheng
keywords: S phase arrest; apoptosis; bakuchiol; breast cancer cell; estrogenic activity; medaka; zebrafish