This study aimed to investigate the anticancer effects of salidroside (SAL), and its inhibitory effects on the proliferation and apoptosis of SKOV3 cells, and its regulatory activities on the carcinogenesis, progression, and growth of ovarian cancer in a mouse model. The expression levels of ARID1A, PTEN, TGF-β, SMAD, FSHR, and ER-β genes and proteins in SKOV3 cells were detected and analyzed. A total of 125 female mice were used to establish ovarian cancer models, which were randomly divided into the control group (CG), Test-1 group, Test-2 group, and Test-3 group. During the experiment, the tumor volume and multiple related indicators in the ascites and serum of mice were dynamically measured. The results showed that the proliferation inhibition rate of SKOV3 cells in the SAL-treated groups was significantly increased as compared with CG. The average daily tumor growth rate of the Test-1, Test-2, and Test-3 groups was lower than that of the CG. On day 50, the p53 gene expression levels in the Test-1, Test-2, and Test-3 groups were increased by 103.77%, 150.94%, and 124.53%, respectively, relative to the CG. The ovarian cancer cells were found in tissue sections at day 14. The contents of CA125 and VEGF-A in the ascites of the SAL-treated groups were higher than those in the C. However, the increments were gradually reduced with the increase in SAL dosage in a concentration-dependent manner. On day 50, the counts of Treg cells in the Test-2 and Test-3 groups were increased by 14.39% and 30.14% compared with the Ctrl group. Additionally, the concentrations of IL-2, IL-10, and TGF-β in the Test-3 group were increased by 271.56%, 663.69% and 557.58%, respectively. In conclusion, this study is the first time to reveal that salidroside can inhibit the proliferation of SKOV3 cells by promoting their apoptosis. It can suppress the carcinogenesis, progression, and growth of ovarian cancer by upregulating the expression of PTEN, ARID1A, TGF-β, and SMAD4, increasing the production of CA125, VEGF-A, IL-2, and IL-10, and elevating the levels of CD3 ⁺ , CD8 ⁺ , and Treg cells in the ascites. SAL at a dose of 400 μg/g exhibited the most potent inhibitory effect on ovarian cancer, which lays a solid foundation for the development of salidroside as a novel anticancer drug.