PEPT1 is a vital member of the proton-dependent oligopeptide transporters family (POTs). Many studies have confirmed that PEPT1 plays a critical role in the absorption of dipeptides, tripeptides, and pseudopeptides in the intestinal tract. In recent years, several studies have found that PEPT1 is highly expressed in malignant tumor tissues and cells. The abnormal expression of PEPT1 in tumors may be closely related to the progress of tumors, and hence, could be considered as a potential molecular biomarker for the diagnosis, treatment, and prognosis in malignant tumors. Furthermore, PEPT1 can be used to mediate the targeted delivery of anti-tumor drugs. Herein, the expression, regulation, and role of PEPT1 in tumors in recent years have been reviewed.