Abstract: Autophagy, a highly conserved lysosome-dependent intracellular degradation pathway in eukaryotic cells, plays an important role in many physiological and pathological processes of cells. With the in-depth analysis of the key genes and signaling pathways of autophagy, the relationship between autophagy and drug toxicity has been gradually revealed. As a class of drugs with severe toxic and side effects, anti-tumor drugs often cause damage to other organs while treating tumors. Autophagy acts as a protective mechanism to reduce tissue damage in toxicity caused by some anti-tumor drugs, however, persistent or excessive autophagy activation can also lead to cell death and induce toxicity. This article reviews the regulatory mechanism as well as the physiological and pathological effects of autophagy, with particular focus on the role and mechanism of autophagy in toxicity caused by anti-tumor drugs, which will improve our comprehensive understanding of autophagy in anti-tumor drug toxicity, and help to discover the key elements for the shifting of the role of autophagy, in the hope of providing new insights and targets for the treatment of anti-tumor drugs-induced toxicity and the development of innovative drugs based on toxicity mechanism.