Lung cancer caused by diverse changes in cells resulted by exposure to carcinogens found in tobacco smoke, the environment, or sequential accumulation of genetic changes to the normal epithelial cells of the lung. An assessment was made of the anti-proliferative activity of constituents from silkworm feces against 11 human cancer cell lines, including A549 and H727 lung cancer cell lines, using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The ethanol extract of silkworm feces was proved to have anti-proliferative activity against all 11 species of human cancer cell lines. The biologically active constituent was characterized as vomifoliol (blumenol A) (1) and stigmasterol (2) by spectroscopic analysis ,including MS and NMR. In conclusion, global efforts to reduce the level of antitcancer agents justify further studies on the silkworm feces-derived materials containing vomifoliol and stigmasterol as potential anticancer products or lead compounds for the prevention or eradication from human lung cancer.