Herein, puromycin treatment induced significant cytotoxicity in p53 wild-type HCT116 cells more than in p53 mutant or null SW620, HCT15 colon cancer cells, and H1299 non-small lung cancer cells by MTT assay, implying potent involvement of the p53 signaling pathway in the antitumor effect of puromycin. Here, TP53 is linked to colonic neoplasm.