A novel derivative of Genistein inhibits proliferation of ovarian cancer HO-8910 cells by regulating reactive oxygen species*

Abstract

ObjectiveTo investigate the anticancer effect of a novel derivative of genistein (5-hydroxy-4’-nitro-7-propionyloxy-genistein, HNPG) on human ovarian cancer HO-8910 cells and its possible molecular mechanism.MethodsHO-8910 cells were cultured in vitro, and the inhibitory effect of HNPG on proliferation was determined using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The effect of HNPG on inducing apoptosis was examined using FCM with Annexin V-FITC and propidium iodide staining. The effect of HNPG on regulating reactive oxygen species (ROS) was measured using FCM with 2’,7’-di chlorodihydro-fluorescein diacetate staining. The effect of HNPG on modulating mitochondrial membrane potential (MMP) was determined using FCM with lipophilic cationic dye 2 (6 Amino 3 imino 3H xanthen 9 yl) benzoic acid methyl ester (Rh123) staining. The bioactivity of superoxide dismutase (SOD) and catalase(CAT) and the content of glutathione (GSH) and malondialdehyde (MDA) were detected using enzyme-linked immunosorbent assay. The related apoptotic proteins, including bcl-2, bax, cyt-c, and cleaved-caspase-3, were assessed using western blotting.ResultsHNPG exhibited dramatic antitumor activity against HO-8910 cells in vitro, inhibited proliferation, and induced apoptosis in a time- and dose-dependent manner. These effects were accompanied by reduced bioactivity of SOD and CAT, reduced GSH content, and enhanced MDA content. Simultaneously, the amount of ROS was increased and the level of MMP was reduced, along with upregulation of mitochondrial apoptosis pathway-related proteins, bax, cyt-c, and cleaved-caspase-3; bcl-2 protein was downregulated.ConclusionHNPG inhibited proliferation of human ovarian cancer HO-8910 cells in vitro, which might be related to decreased bioactivity of SOD and CAT. HNPG also reduced GSH content, which resulted in ROS accumulation in cells, damaged the integrity of mitochondrial membrane, and induced cell apoptosis.