Mechanism of ergosterol peroxide on MCF-7 breast cancer cells based on network pharmacology and in vitro experiments
This study investigated the effects of ergosterol peroxide(EP)on the proliferation and apoptosis of MCF-7 breast cancer cells,explored its possible mechanisms of action,and verified the effects and mechanisms by in vitro experiments.Network pharmaco-logy was used to screen the target proteins of EP and construct target networks and protein-protein interaction(PPI)networks to predict the potential target proteins and related pathways involved in EP anti-breast cancer effects.The MTT assay was performed to measure the inhibitory effect of EP on MCF-7 cell proliferation,and the colony formation assay was used to assess the cell cloning ability.Flow cytometry and laser confocal microscopy were employed to evaluate cell apoptosis,mitochondrial membrane potential and reactive oxy-gen species(ROS)levels.Western blot analysis was conducted to examine the expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cytochrome C(Cyt C),caspase-7,cleaved caspase-7,phosphatidylinositol 3-kinase(PI3K),and se-rine/threonine kinase B(AKT)in MCF-7 cells treated with EP.The results of network pharmacology prediction yielded 173 common targets between EP and breast cancer;the results of Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis showed that EP treatment for breast cancer mainly affected the signaling pathways such as cancer pathway,PI3K-AKT signaling pathway,cel-lular senescence signaling pathway,and viral carcinogenesis pathway;and the MTT assay results showed that the viability of MCF-7 cells in the EP group was significantly lower than that in the control group,exhibiting a time-and concentration-dependent trend,and EP can inhibit colony formation of MCF-7 breast cancer cells.Treatment with 10,20,and 40 μmol·L-1 EP for 24 h resulted in a sig-nificant increase in the total apoptosis rate of MCF-7 cells,a significant decrease in mitochondrial membrane potential,and a signifi-cant increase in ROS levels.In addition,treatment with EP led to an upregulation of Cyt C,Bax,and cleaved caspase-7 protein ex-pression,and a downregulation of p-PI3K,p-AKT,and Bcl-2 protein expression in MCF-7 cells.Studies have shown that EP inhibits MCF-7 breast cancer cell proliferation and reduces colony formation by a mechanism that may be related to the PI3K-AKT pathway me-diating the mitochondrial apoptotic pathway.
ergosterol peroxidebreast cancerMCF-7 cellsnetwork pharmacologycell apoptosismitochondrial membrane po-tentialreactive oxygen species
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