Background The LIM-1863 Cell Line is one of the colon cancer types considered to be responsible for a high rate of deaths, and the glycoalkaloids being natural substances existing in the Solanum species have anticancer effects. Objective This research aims at studying the effect of the glycoalkaloids on viability of the LIM-1863 cancerous cells in-vitro. Materials and Methods The glycoalkaloids in this study are extracted by the ultrasonic waves technique and detecting them by the Thin Layer Chromatography (TLC) in addition to incubating of the LIM-1863 cells with different concentrations of the glycoalkaloids for 48 hours and then assessing of the cell viability using the MTT assay.Results The findings showed that the glycoalkaloids have a toxic effect on the LIM-1863 cells and that half of the inhibiting concentration (IC 50 ) of the Solanum fruits extract: (Solanum nigrum L.), (Solanum villosum Mill.) and (Solanum elaeagnifolium Cav.) on the LIM-1863 cells have the (164.7, 35.91 and 12.14 µg/ml) values successively. Conclusion The observations indicated that the glycoalkaloids are able to inhibit the colon cancer cell proliferation.