The aim of the present study was to perform comparative genomics using gene expression profile datasets of mice and humans who had been treated with anticancer drugs, to determine the similarities and differences in the antitumor mechanisms in the two mammals. This involved data mining of antitumor gene expression regulation, and screening of genetic loci from experimental mouse models of antitumor targets, to provide a theoretical basis of drug design. Subsequently, 9 overlapping genes with opposite expression patterns were identified across mouse and human cell lines that were treated with a specific cyclin‑dependent kinase 4/6 inhibitor, PD0332991. These genes included LIM homeobox 2, adenomedullin, bone marrow stromal cell antigen 1, caveolin 1, histone cluster 1 (HIST1) H2B family member C, HIST1 H3 family member F, low density lipoprotein‑receptor related protein 11, prolyl 4‑hydroxylase subunit α1 and torsin family 3 member A. In addition, the janus kinase‑signal transducer and activator of transcription signaling pathway, Toll‑like receptor signaling pathway, T cell receptor signaling pathway and the nucleotide‑binding oligomerization domain‑like receptor signaling pathway were identified as candidate pathways for explaining antitumor mechanisms.