Abstract OBJECTIVE To explore the intervention mechanism of Guizhi Fuling pills on breast cancer based on multi-data platform and bioinformation technology, and to verify the analysis results by cell test. METHODS The data set related to breast cancer was downloaded from GEO database to analyze the differential genes of breast cancer. The main active components and target genes of Guizhi Fuling pills were screened from TCMSP database. The key target genes of Guizhi Fuling pills in the treatment of breast cancer were mapped by the two groups of target genes. TCGA database was used to analyze the expression of key target genes in breast cancer. Through TIMER, CPTAC Data Portal, and Kaplan-Meier plotter, TISIDB, SangerBox and other data platforms, the relationships of the key genes expression and immune micro-environment, immune infiltration level, genomic heterogeneity, immune subtypes, molecular subtype, and poor prognosis were analyzed in breast cancer. KEGG pathway enrichment of key genes was performed searching for possible signaling pathways by Metascape analysis platform. Finally, CCK-8 assay, cell scratch assay, Transwell chamber assay and Western blotting technique were used to verify the results in vitro. RESULTS There were 42 active components and 185 targets in Guizhi Fuling pills, and 14 key targets related to the treatment of breast cancer were mapped with GEO database. Six key target genes for the treatment of breast cancer were obtained by comparison with breast cancer differential genes in TCGA. The expression of these 6 key genes was strongly correlated with the levels of 6 immune cells, 3 immune microenvironment scores, immune subtypes and molecular subtypes in breast cancer(P<0.05), also showed consistency in the correlation of genomic heterogeneity, and was closely associated with poor prognosis(P<0.05). KEGG enrichment analysis showed that these 6 genes were mainly enriched in PI3K/Akt signaling pathways. Cell test showed that Guizhi Fuling pills could significantly inhibit the proliferation, migration and invasion of HCC1937 cells, and also reduce the relative expression of p-Akt/Akt and p-PI3K/PI3K proteins in HCC1937 cells in a dose-dependent manner. CONCLUSION Guizhi Fuling pills may interfere with breast cancer by blocking PI3K/Akt signaling pathway and regulating the tumor immune microenvironment.