基于生信分析探索FOSL1对肝癌索拉非尼治疗抵抗的影响

Exploring the effect of FOSL1 on sorafenib resistance in liver cancer based on bioinformatics analysis

目的:探究FOS样抗原1(FOS-like antigen 1,FOSL1)在肝癌组织中的表达及其对索拉非尼治疗抵抗的影响。方法:从公共数据库中获取数据进行WGCNA分析,对获取的差异表达基因(differentially expressed genes,DEGs)进行GO功能富集和KEGG通路富集分析。选取血管生成通路差异表达基因,采用STRING数据库建立PPI网络,使用MCODE插件提取排名前7位的关键基因。在人类蛋白质图谱网站对这7个关键基因在肝癌血管内皮细胞表达情况进行筛选。采用Western blot法检测人微血管内皮细胞-1(human microvascular endothelial cells-1,HMEC-1)及肝癌血管内皮细胞(tumor-derived endothelial cells,TEC)中FOSL1的表达,CCK-8法检测HMEC-1及TEC细胞对索拉非尼敏感性,免疫组化检测肿瘤组织及正常肝组织中FOSL1表达。结果:GO功能富集分析图和KEGG通路富集分析图显示“索拉非尼抵抗”和“索拉非尼不抵抗”患者DEGs主要富集在管腔形成和血管生成等通路,KEGG通路主要富集在Wnt信号通路和VEGF信号通路。WGCNA分析得到关键模块,通过对关键模块基因筛选,最终得到JUND、JUNB、IL17RC、IL17RA、IL17F、IL1B、FOSL17个关键基因,其中FOSL1在血管生成中发挥重要作用。通过人类蛋白质图谱网站,发现FOSL1在肝脏各类细胞中表达,但在内皮细胞中表达较高。Western blot检测显示,FOSL1在TEC细胞中的表达水平明显高于HMEC-1细胞。免疫组化染色发现,肿瘤组织中FOSL1高表达,正常肝组织中FOSL1表达水平较低。CCK-8法检测显示,HMEC-1细胞对索拉非尼较敏感,而TEC细胞对索拉非尼不敏感。结论:FOSL1在肝癌血管内皮细胞中高表达,可能与促进血管内皮细胞增殖及索拉非尼治疗抵抗相关。

Objective:To investigate the expression of FOS-like antigen 1(FOSL1)in liver cancer and the effect of FOSL1 on sorafenib resistance.Methods:Obtain data from public databases for WGCNA analysis,and perform GO functional enrichment and KEGG pathway enrichment analysis on the differentially expressed genes(DEGs)obtained.Select differentially expressed genes in the angiogenesis pathway,establish a PPI network using the STRING database,and extract the top 7 key genes using the MCODE plugin.Screen the expression of these 7 key genes in liver cancer endothelial cells on the Human Protein Atlas website.Western blot was used to detect the expression of FOSL1 in human microvascular endothelial cell-1(HMEC-1)and tumor-derived endothelial cells(TEC),CCK-8 method was used to detect the sensitivity of HMEC-1 and TEC cells to sorafenib,and immunohistochemistry was used to detect the expression of FOSL1 in tumor tissue and normal liver tissue.Results:GO functional enrichment analysis and KEGG pathway enrichment analysis showed that DEGs in patients with“sorafenib resistance”and“sorafenib non resistance”were mainly enriched in luminal formation and angiogenesis pathways,while KEGG pathway was mainly enriched in Wnt and VEGF signaling pathways.WGCNA analysis identified key modules,and ultimately identified JUND,JUNB,IL17RC,IL17RA,IL17F,IL1B,and FOSL17 key genes through screening of key module genes,among which FOSL1 played an important role in angiogenesis.It was found that FOSL1 was expressed in various liver cells through the Human Protein Atlas website,but more higher in endothelial cells.Western blot analysis showed that the expression level of FOSL1 in TEC cells was significantly higher than that in HMEC-1 cells.Immunohistochemical staining revealed high expression of FOSL1 in tumor tissue and low expression in normal liver tissue.CCK-8 assay showed that HMEC-1 cells were sensitive to sorafenib,while TEC cells were not sensitive to sorafenib.Conclusion:FOSL1 is highly expressed in liver cancer endothelial cells,which may be associated with promoting endothelial cell proliferation and sorafenib resistance.