A method for constructing a subgroup B recombinant human adenovirus vector Ad11-5EP. The method includes substituting a 365 bp fragment including an enhancer and a promoter of an upstream coding sequence of Ad5 E1A for a corresponding region of a serotype Ad11 of the subgroup B human adenovirus vector by homologous recombination to construct the subgroup B recombinant human adenovirus vector Ad11-5EP. A subgroup B recombinant human adenovirus vector Ad11-5EP constructed by the method and the use thereof for treatment of tumors are also provided.