Sambucus ebulus is a native botany and exists in large amount in Iran and consists of anti-cancer substances such as ebulin (RIP-II, ribosome inactivated protein-II), flavonoids, etc. Isolation and identification of some potent anti-tumor compounds from medicinal plants, has motivated researchers to screen different parts of plant species for anti-tumor effects. In previous studies, anti-inflammatory effects of n-hexane and methanolic extracts of S. ebulus , nephro- and hepato-toxic effects of ethylacetate extract of this plant and evaluation of role of vitamins C and E on prevention of cellular and pathological disorders induced by the ethyl acetate extract was performed and reported. So, cytotoxic activity and IC50 of specific concentrations of ethyl acetate extract of fruits of S.ebulus on 4 normal and cancer cell lines was studied. Also, Etoposide, a chemotherapeutic drug, was administered to control positive group. The normal cell lines were CHO and rat fibroblast and cancer cell lines were HepG2 and CT26. The cytotoxic effects and IC50 of the extract on the cell lines were studied followed by MTT assay. The results showed that the ethyl acetate extract of Sambucus ebulus possesses lower IC50 in the cancer cell lines in comparison with the normal cell lines. On the other hand, the extract possesses higher IC 50 in comparison with Etoposide on all 4 normal and cancer cell lines (P<0.05), but it manifested a good cytotoxic compound which can introduce as an anticancer compound.