Background: Colon cancer is a major health problem with increasing mortality rates worldwide. Aim: Is to evaluate the ability of Ferula Assafoetida (F. assafoetida) to induce differentiation of colon cancer cells to function as normal cells. Methods: The cytotoxic effect of F. assafoetida was assessed against Caco cells using the MTT assay. Cell cycle analysis and apoptosis were assessed using CytellTM cell system. The total antioxidant (TA), glutathione (GSH), Malondialdehyde (MDA) concentrations, and alkaline phosphatase (ALP) activity were also assessed. The JNK and MAPK expressions were evaluated using qRt-PCR. Results: There was a significant increase in the cell number treated with F. assafoetida (53.85±0.03%), and those treated with NaBT (54.6±0.10%) in sub-G1 phase, compared to the untreated cells (0.78±0.03%, p<0.001). Apoptosis was significantly increased in the Caco cells treated with F. assafoetida (49.1± 0.14 %) compared to those treated with NaBT (27.3±0.10 %, p<0.001), and untreated cells (11.1±0.02 %, p<0.001). DNA degradation was observed in Caco cells treated with F. assafoetida at a dose-dependent manner, where complete degradation occurred at the dose of IC50 (342.9 μg/mL). F. assafoetida induced a significant increase in the TA concentration and GSH, while a significant decrease in the MDA levels (p<0.001, for all). Also, there was a significant increase in ALP activity in Caco cells (0.53±0.26 U/mL) compared to the control cells (0.05±0.02 U/mL, p=0.045). There was a significant upregulation of JNK and MAPK expression in Caco cells treated with F. assafoetida compared to the controls [2.59±0.01 (p<0.001), and 3.62±0.01 (p<0.001); respectively]. Conclusion: F. assafoetida is a potentially successful therapeutic and differentiating agent for colon cancer.