Abstract: Objective: To explore the effect of raloxifene(RAL) on the malignant phenotypes of oral squamous cell carcinoma(OSCC) and its potential molecular mechanisms. Methods: The target genes of RAL were predicted using TargetNet, SEA, and STITCH databases. The OSCC-related target genes were screened through GeneCards, MalaCards, and OMIM databases. The intersection targets were obtained using Venny. The interaction targets were analyzed for GO and KEGG enrichment using Metascape. The core targets were screened using CytoNCA, Cytohubba, and MCODE plugins. The half-maximal inhibitory concentration(IC50) and proliferation ability of cells were detected by cell counting kit-8(CCK-8) assay. The mRNA levels of core targets were detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR). The clone formation, migration, and invasion abilities of cells were detected by clone formation, scratch, and Transwell chamber assays. The apoptosis and cell cycle of cells were detected by flow cytometry. The differentially expressed genes and the potential molecular mechanisms were screened by RNA sequencing. The activation levels of related signaling pathways were detected by western blotting. Functional recovery experiments were conducted using the PI3K/AKT pathway activator Recilisib, and the in vivo efficacy was evaluated through a nude mouse xenograft tumor model. Results: A total of 58 potential targets of RAL in the treatment of OSCC were screened out, among which SRC, CASP3, HSP90 AA1, TNF, and AKT1 were identified as core targets. RAL increased the mRNA levels of SRC and CASP3 and decreased those of HSP90AA1 and TNF(all P<0.05). RAL inhibited cell proliferation, clone formation, migration, and invasion, promoted cell apoptosis in a dose-dependent manner, and arrested the cell cycle at the G0/G1 phase(all P<0.05). Combined network pharmacology and RNA sequencing data revealed that the PI3K/AKT signaling pathway was the core functional pathway of RAL in combating OSCC. RAL treatment led to downregulation of p-PI3K and p-AKT phosphorylation levels(all P<0.05). PI3K/AKT pathway activators could significantly reverse the inhibitory effects of RAL on the malignant phenotype of OSCC cells(all P<0.01), but failed to reverse its regulation on core targets such as SRC; in vivo animal experiments confirmed that RAL could significantly inhibit the growth of xenograft tumors, with an inhibition rate of 56%(all P<0.01). Conclusion: RAL may inhibit the activation of the PI3K/AKT signaling pathway and independently regulate upstream targets, thereby suppressing the proliferation, colony formation, migration and invasion abilities of OSCC cells, promoting apoptosis and blocking the cell cycle.