青蒿琥酯通过FABP5调节PI3K/AKT通路影响肝癌细胞的增殖和迁移

Artesunate affects the proliferation and migration of HCC cells by regulating the PI3K/AKT pathway through FABP5

目的基于网络药理学与生物信息学探寻青蒿琥酯(ART)治疗肝细胞癌(HCC)的潜在靶点、通路及相关机制。方法通过PharmMapper数据库筛选ART的潜在作用靶点,构建PPI网络并进行可视化分析。从TCGA数据库下载424例HCC样本数据,筛选潜在靶点基因的数据信息,并分析差异表达的靶点基因;利用单-多因素COX回归分析,筛选关键靶点基因;用分子对接软件对ART和关键靶点基因进行分子对接;用网络数据库GEPIA2对关键靶点基因进行差异表达和生存分析;通过R语言进行KEGG通路富集分析;通过MTS实验、划痕实验验证ART对肝癌细胞增殖和迁移的影响;qPCR和Western blot验证FABP5在肝癌细胞中的表达,Western blot验证PI3K/AKT通路蛋白变化水平。结果通过网络药理学筛选出282个潜在靶点基因,最终筛选出3个关键靶点基因;FABP5在HCC中的生存分析有显著的统计学意义(P<0.01);分子对接显示ART和FABP5的结合能最高;通路富集分析表明FABP5主要富集在PI3K/AKT信号通路;细胞实验验证ART能抑制FABP5的表达水平,ART可以调节PI3K/AKT通路,ART能抑制肝癌细胞的增殖和迁移。结论ART可能通过抑制FABP5调节PI3K/AKT通路进而影响肝癌细胞的增殖和迁移,FABP5可能作为ART治疗HCC的新靶点基因。

Objective To explore the potential targets,pathways and related mechanisms of artesunate(ART)in the treatment of hepatocellular carcinoma(HCC)based on network pharmacology and Bioinformatics.Methods The potential targets of ART were screened through pharmmapper database,PPI network was constructed and visual analysis was performed.The data of 424 HCC samples were downloaded from TCGA database,the data information of potential target genes were screened,and the differentially expressed target genes were analyzed.The key target genes were screened by univariate-multivariate COX regression analysis.Molecular docking software was used to conduct molecular docking between ART and key target genes.Differential expression and survival analysis of key target genes were performed using network database GEPIA2.KEGG pathway enrichment analysis was performed by R language.The effects of ART on the proliferation and migration of HCC cells were verified by MTS assay and scratch test.The expression of FABP5 in HCC cells was verified by qPCR and Western blot.Western blot was used to verify the protein changes of PI3K/AKT pathway.Results A total of 282 potential target genes were screened by network pharmacology.Finally,three key target genes were screened out.The survival analysis of FABP5 in HCC was statistically significant(P<0.01).Molecular docking showed that ART and FABP5 had the highest binding energy.Pathway enrichment analysis showed that FABP5 was mainly enriched in PI3K/AKT signaling pathway.Cell experiments verified that ART could inhibit the expression level of FABP5,that ART could regulate the PI3K/AKT pathway,and that ART could inhibit the proliferation and migration of HCC cells.Conclusion FABP5 may regulate the PI3K/AKT pathway by inhibiting FABP5 and thus affect HCC cell proliferation and migration,and FABP5 may serve as a new target gene for ART therapy of HCC.