Abstract: Objective To investigate the effect of sevoflurane (Sev) on the malignant biological behaviors of ovarian cancer cells and its possible mechanism. Methods Human endometrial epithelial cells (hEECs) and human ovarian cancer cell line (SKOV3) were treated with sevoflurane at the concentrations of 1%, 2%, and 4% (V/V). The cell proliferation was measured by MTT assay. SKOV3 cells were randomly divided into five groups: control group, Sev low concentration (1%) group, Sev medium concentration (2%) group, Sev high concentration (4%) group, and Sev high concentration + lipopolysaccharide (1 mg/L) group. Cell migration and invasion abilities were assessed by Transwell assays. Cell apoptosis was detected by flow cytometry. Inflammatory cytokine levels were measured by ELISA. The expression of genes related to proliferation, apoptosis, and invasion were analyzed by quantitative real-time PCR (qRT-PCR). The expression of proteins involved in proliferation, apoptosis, invasion, and the TLR4/MyD88/TRAF6 signaling pathway were detected by Western blot. Results Treatment with 1%, 2%, and 4% sevoflurane gradually decreased the survival rate of SKOV3 cells in a dose-dependent manner, with no effect on the survival of hEECs. Additionally, treatment with 1%, 2%, and 4% sevoflurane reduced the invasion and migration rates of SKOV3 cells and promoted apoptosis. It also decreased the levels of tumor necrosis factor-α, interleukin-6, and interleukin-1β in the supernatant, and inhibited the expression of TLR4, MyD88, and TRAF6 proteins (P<0.01). Co-treatment with 4% sevoflurane and lipopolysaccharide reversed the effects of sevoflurane on SKOV3 cells. Conclusions Sevoflurane may regulate the TLR4/MyD88/TRAF6 signaling pathway to inhibit the proliferation, migration, and invasion of ovarian cancer cells, and promote apoptosis in a dose-dependent manner.