Effects of Herba Houttuyniae on the biological behavior of liver cancer cells and a network pharmacology analysis
Objective To investigate the effects of Herba Houttuyniae on the biological behavior of liver cancer cells and related potential molecular mechanisms.Methods Liver cancer cells SMMC-7721 and SK-Hep-1 were treated with different concentrations of Herba Houttuyniae extract.Cell viability was detected by CCK-8.Transwell assays were used to evaluate the migration and invasion ability of the liver cancer cells.The expression of the metastatic-related protein MMP-9 was detected by cellular immunofluorescence assay.The reactive oxygen species(ROS)levels in the cells were measured by using ROS detection kit,and cell apoptosis was assessed by TUNEL staining.The expression levels of apoptosis-related proteins were analyzed by Western blot.Network pharmacology was employed to screen the active components of Herba Houttuyniae and to construct a protein-protein interaction(PPI)network to identify the therapeutic targets of Herba Houttuyniae in liver cancer.GO and KEGG pathway enrichment analyses were performed to explore the signaling pathways associated with the target of Herba Houttuyniae in anti-liver cancer.The qRT-PCR was used to detect the mRNA expression levels of key targets.Results The CCK-8,Transwell,and cellular immunofluorescence assays indicated that Herba Houttuyniae extracts inhibited the viability,migration,and invasion abilities of SMMC-7721 and SK-Hep-1 cells,as well as the expression of the metastasis-related protein MMP-9(all P＜0.05).TUNEL staining,ROS detection,and Western blot results showed that Herba Houttuyniae extracts promoted apoptosis in SMMC-7721 and SK-Hep-1 cells,elevated intracellular ROS levels,and increased the expression of the pro-apoptotic protein BAX(all P＜0.05).Network pharmacology predicted that Herba Houttuyniae treated liver cancer primarily by targeting TNF,AKT1,TP53,etc.involving cancer,TNF,PI3K-AKT and other signaling pathways.The qRT-PCR confirmed that Herba Houttuyniae extracts decreased the mRNA expression levels of TNF-α and AKT1 genes,while increased the mRNA expression level of the TP53 gene in SMMC-7721 and SK-Hep-1 cells(all P＜0.05).Conclusions Herba Houttuyniae may inhibit the viability,migration,and invasion abili-ties of liver cancer cells and promote the apoptosis by regulating genes such as TNF,AKT1,and TP53.
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