EpCAM (Epithelial Cell Adhesion Molecule) is a cell surface glycoprotein. Concurrently, its cleavage fragments, EpEX (extracellular domain) and EpICD (intracellular domain), exert bidirectional regulation on ERK (Extracellular Signal-Regulated Kinase) phosphorylation. Specifically, EpEX binds to EGFR (Epidermal Growth Factor Receptor) to promote ERK phosphorylation, while EpICD interacts with PKC (Protein kinase C) to inhibit this pathway, consequently influencing cell motility and proliferation. This study aims to elucidate the dependence and regulatory roles of EpCAM on different signaling pathways during its modulation of ERK phosphorylation and cell migration. Using CaCO2 cells as the model, the release of EpEX and EpICD was selectively inhibited, and the cells were stimulated with EGF (Epidermal Growth Factor). Signal protein expression, cell migration, and proliferation were analyzed using Western blot, Wound healing assay, and Proliferation assay, respectively. The results showed that concerning ERK phosphorylation, inhibiting EpICD generation (via DAPT treatment) relieved the PKC pathway's inhibition on ERK, leading to the most significant enhancement in ERK phosphorylation activity. Conversely, inhibiting EpEX generation (via TAPI-1 treatment) attenuated the EGFR downstream signaling, resulting in the most pronounced inhibitory effect on cell proliferation and migration. In summary, EpCAM simultaneously exerts bidirectional regulation on ERK activity through its two cleavage fragments, EpEX and EpICD, under EGF stimulation, thereby impacting cell growth and migration. These findings establish an important foundation for investigating tumor cell invasiveness and signal transduction regulation. Future studies employing fluorescence microscopy and multiplex protein analysis may further clarify the potential involvement of EpCAM in cancer metastasis and therapeutic strategies.