Mechanism of rapamycin combined with flagellin inhibiting 4T1 breast cancer cells in vitro based on mRNA high-throughput sequencing.

AIM: This study explores how the combination of rapamycin (Rapa) and flagellin (FliC) affects the inhibition of 4T1 breast cancer cells. The approach involves using mRNA high-throughput sequencing to examine the underlying mechanisms of this combination therapy in vitro. METHODS: 4T1 breast cancer cells were divided into four groups: control group, Rapa group, FliC group, and Rapa+FliC group. The changes in cell viability and apoptosis were detected by the CCK-8 method and flow cytometry. Concurrently, the KEGG pathway of differentially expressed genes (DEGs) was analyzed by high-throughput mRNA sequencing. Furthermore, the DEGs between the Rapa+FliC group and Rapa groups were analyzed using STRING. The PPI network of DEGs was then constructed, and the Hub genes were subsequently screened. The protein expression of the Hub gene was verified based on the HPA database. RESULTS: CCK-8 assays and flow cytometry analysis revealed that the combination of Rapa and FliC significantly increased both the inhibition and apoptosis rates in 4T1 breast cancer cells compared to the rates observed with Rapa or FliC alone (P<0. 05). Transcriptome sequencing indicated 579 DEGs between the Rapa group and the control group, predominantly in the PI3K/ Akt signaling pathway. In contrast, DEGs between the FliC group and control were mainly concentrated in signaling pathways like NOD-like receptor signaling. Additionally, 150 DEGs were identified between the Rapa+FliC group and the Rapa group, focusing primarily on pathways such as mTOR. From the protein-protein interaction (PPI) network, ten hub genes, including Atm and Itga2, were identified. CONCLUSION: The combination of Rapa+FliC could inhibit the viability of 4T1 breast cancer cells in vitro and promote apoptosis, potentially through the PI3K/Akt/mTOR signaling pathway. The genes Atm and Itga2 could be pivotal in mediating the joint effect of this combination therapy. [ABSTRACT FROM AUTHOR]