Urolithin A (UA) emerges as a promising natural anticancer agent and adjunct in treating colorectal cancer (CRC). This study aims to delve into the mechanism of action of UA in CRC cells. In this investigation, two colon cancer cell lines, namely HT29 and SW620, underwent treatment with specific cellular apoptotic inhibitors preceded by UA pre-treatment. The evaluation of UA's mechanism in CRC cells involved flow cytometry and western blotting. The findings revealed that apoptotic inhibitors U0126 and LY294002 mitigated the apoptotic effect of UA across all cell lines. Furthermore, in HT29, elevated levels of p-ERK and p-AMPK were observed. In SW620, the expression levels of phosphorylated AKT and the cellular proliferation regulator mTOR decreased significantly (P < 0.001), while the levels of the tumor suppressors, p-c-RAF and p-PTEN, increased. These observations suggest that UA targets the p-c-RAF/MEK/p-ERK signaling pathway in the early stages of CRC, while its mechanism in metastasis is contingent on the PI3K/p-AKT/mTOR pathway, as well. This study presents the initial evidence elucidating UA's anticancer properties by influencing these signaling pathways during both the early stages and metastasis of CRC.