Abstract: Methotrexate is an anti-folate antitumor drug, which is often used to treat cancer, rheumatoid arthritis and psoriasis. Methotrexate has a significant therapeutic effect, but its treatment window is narrow. Various transporters are involved in the disposal of methotrexate in vivo. When methotrexate is used in combination with other drugs, drug interactions based on transporters are likely to occur, leading to side effects or changes in efficacy. This article summarized the transport-mediated drug-drug innteractions of methotrexate to provide references information for rational administration of medication in clinic.