In this study, we demonstrate that L3 expression in colon tumor tissues is downregulated; L3 mRNA decrease associated with malignance progression and tumor grade and was inversely proportional to the ratio Bcl-2/Bax; L3 overexpression in 5-FU treated colon cancer cells decreased clonogenic potency, cell migration and cell viability, and stimulated apoptotic cell death by inducing late apoptosis. Here, RPL3 is linked to colonic neoplasm.