Provided are a recombinant herpes simplex virus, a preparation method and use thereof. The recombinant herpes simplex virus comprises a vector and foreign genes encoding at least two cytokines, wherein the vector is a herpes simplex virus with genes encoding ICP34.5 and ICP47 deleted, and optionally with at least one of genes encoding ICP6, TK and UNG deleted, and the insertion site/sites of the foreign genes is in at least one of the positions where the genes encoding ICP34.5, ICP47, ICP6, TK and UNG are deleted in the vector.