Objective To investigate the effects of Pinella’s ingredients on the viability of cells, morphology, microstructure, cell cycle and apoptosis in human cervical cancer cell lines. Methods The β-sitosterol and/or total protein of Pinella were incubated at different concentrations with cervical cancer cell line SiHa. The effects of β-sitosterol and/or total protein of Pinella on the viability of cells were tested by MTT assay. The effects of β-sitosterol on morphology, microstructure, cell cycle and apoptosis were studied by phase-contrast microscope, electron microscope and flowcytometry, respectively. Results β-sitosterol could obviously inhibit the viability of SiHa cells in a time- and dose-dependent manner. The total protein of Pinella had no effects on SiHa cells’ viability. 20 μmol/L β-sitosterol induced the accumulation of SiHa cells in S phase in the cell cycle. And the percents of apoptosis and necrosis increased. The morphology and microstructure of SiHa cells changed significantly after treated with 20 μmol/L β-sitosterol. Conclusions The total protein of Pinella had little influence on the viability of cervical cancer cells SiHa. The viability of SiHa cells could be suppressed by β-sitosterol. β-sitosterol could induce the accumulation of cells in S phase and the percent of apoptosis and necrosis. The morphology and microstructure changed significantly after treated with β-sitosterol. Therefore, β-sitosterol might be a prospect safe and low toxicity anti-cervical cancer drug.