〔Abstract〕 Objective To explore the activity of auranofin against ovarian cancer cells and its possible molecular mechanism. Methods The dose-response survival curve and IC50of auranofin on ovarian cancer cell lines, SKOV3, Caov3 and SW626 cells and immortalized normal human embryonic kidney HEK-293T cells were determined by CCK-8 method. Cell cycle was determined by flow cytometry. The levels of total glutathione(GSH), reduced GSH and glutathione disulfide(GSSG), thioredoxin reductase(TrxR) and reactive oxygen species(ROS) in cells were determined by microplate reader, and the reduced GSH/GSSG ratio was calculated. Western blot was used to determine the expression of cyclin dependent kinases(CDK)4, CDK6, Cyclin D1, P53, p-P53 and MDM2 in SKOV3 and Caov3 ovarian cancer cells. Results Compared with HEK-293T cells, the dose-response survival curves and IC50values of SKOV3, Caov3 and SW626 cells showed that ovarian cancer cells were more sensitive to auranofin(P<0.05). After SKOV3 and Caov3 cells were treated with the dose of respective IC50concentrations of auranofin, compared with the untreated cells group, the Auranofin IC50group cells′ intracellular levels of GSH, the ratio of reduced GSH/GSSG and the activity of TrxR decreased(t=25.11/31.18, 14.72/19.92, 43.30/10.74, allP<0.05), and the levels of ROS increased(t=23.82/27.71,P<0.05); cells number at G0/G1phases increased, with cells number at S and G2phases decreased(P<0.05); and the expression levels of cell cycle-related proteins CDK4, CDK6, Cyclin D1 and the P53-specific E3 ubiquitin ligase MDM2 were down-regulated(t=7.51/15.59, 17.32/11.26, 20.78/20.78, 24.25/17.32, allP<0.05), while the expression levels of P53 and p-P53 were up-regulated(t=17.32/24.25,12.12/10.39,allP<0.05). Conclusion Auranofin causes oxidative stress in ovarian cancer cells by inhibiting TrxR activity, and by partially degrading MDM2 to stabilize and activate P53, so as to block the cancer cells in G0/G1phase, and exert anti-ovarian cancer activities.