Some novel B-norcholesterols with different substituted groups were synthesized. The antiproliferative activity of the compounds against cervical carcinoma (HeLa), liver cancer (Bel 7404) and gastric cancer (SGC 7901) cells was assayed. The results revealed that the presence of a 6-alkylthiosemicarbazone or 6-cyano group could enhance the antiproliferative activity of these compounds. The induction of compounds 6 and 9 to cancer cell apoptosis were assayed by flow cytometry, and the results showed that the compounds were able to effectively induce cancer cell apoptosis. The research provided a theoretical reference for the exploration of new anti-cancer drug. The results suggest that compounds 6 and 9 based on its abeo-cholestane may constitute a novel class of antiproliferative agents, which deserve further study.