The objective is to study the extraction and refinement processes of Rhizoma Curcumae oil and its inhibitory effect on cervical cancer. Orthogonal test, MTT assay and flow cytometry are used, respectively, to optimize the extraction process of Rhizoma Curcumae oil, and to study its inhibitory effect on CASKI cells. Orthogonal test results show that favorable extraction and refinement effects can be achieved if Rhizoma Curcumae is crushed, passed through an 80-mesh sieve, added with 30-fold amount of water, and steam distilled for 2 h. In the industrial production, the above conditions can easily be met, which can minimize production costs as well. MTT assay and flow cytometry results show that the medium- and high-dose (100, 150 μg/ml) Rhizoma Curcumae oil groups have significantly different growth inhibition rates against CASKI cells, indicating that Rhizoma Curcumae oil can inhibit CASKI cell growth to varying degrees, and in a somewhat dose-dependent manner under the experimental conditions. Rhizoma Curcumae oil can arrest CASKI cell cycle at the G2/M phase.