靶向EGR1对泌乳素瘤的潜在治疗作用

Targeting EGR1 showed potential therapeutic effects on prolactinoma

目的:从基因分子水平出发,探究泌乳素瘤(prolactinoma,PRL)患者差异表达基因,从而探究疾病发展的核心驱动基因。旨在为泌乳素瘤患者的临床诊断、治疗及预后提供潜在的靶标。同时针对核心驱动基因,预测治疗泌乳素瘤患者的分子靶向药物。方法:从GEO(gene expression omnibus)数据库获取泌乳素瘤患者基因表达谱数据,进行泌乳素瘤组织和正常组织差异表达基因(differential expressed genes,DEG)筛选。同时进行加权基因共表达网络分析(weighted gene co-expression network analysis,WGCNA)、蛋白质相互作用网络分析(protein-protein interation,PPI),确定泌乳素瘤发生发展的核心驱动基因。同时进行GO(gene ontology)分析、KEGG(kyoto encyclopedia of genes and genomes)分析及GSEA(gene set enrichment analysis)分析,探究泌乳素瘤患者发生改变的生物功能和信号通路,探究肿瘤发生发展的分子机制。此外,进行体外划痕实验、克隆形成测定、CCK-8实验及流式细胞术实验,验证分子靶向药物的潜在治疗效果。结果:共鉴定出1201个差异表达基因,其中EGR1、MAPK1、MYC、BCL2和CALM1居于重要地位;GO和KEGG分析结果表明,泌乳素瘤的形成主要与纺锤极发生改变、卵母细胞减数分裂受到影响相关;泌乳素瘤很可能与帕金森病存在一定的同源性。同时进行CCK-8测定、克隆形成测定和体外划痕测定,验证了EGR1激动剂Genipin具有潜在抗泌乳素瘤的作用。流式细胞术分析结果显示,随着Genipin药物剂量增加,肿瘤细胞的凋亡百分比增加。结论:EGR1、MAPK1、MYC、BCL2和CALM1是泌乳素瘤的核心驱动基因,对泌乳素瘤的发生发展具有重要影响。EGR1的激动剂Genipin能够在体外抑制泌乳素瘤细胞的增殖和迁移,Genipin对泌乳素瘤具有潜在治疗作用。同时,本研究筛选出了泌乳素瘤发生的高风险信号通路,为其临床诊治提供了新的靶标,具有重要意义。

Objective:To explore the differentially expressed genes in patients with prolactinoma(PRL)from the molecular level of genes,for studying the core driving gene of disease development;to provide potential targets for clinical diagnosis,treatment and prognosis in patients with RPL and predict molecular targeted drugs for treating patients with RPL by focusing on core driving genes.Methods:Gene expression profile data of PRL patients were obtained from the gene expression omnibus(GEO)database,and the differentially expressed genes(DEG)of PRL tissues and normal tissues were screened.At the same time,weighted gene co-expression network analysis(WGCNA)and protein-interaction analysis(PPI)were used to determine the core driver genes of PRL development.At the same time,gene ontology(GO)analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis and gene set enrichment analysis(GSEA)were conducted to explore the altered biological functions and signaling pathways in PRL patients,and to explore the molecular mechanism of tumor development.In addition,in vitro wound-healing test,clone formation assays,CCK-8 assays and flow cytometry experiments were performed to validate potential therapeutic effects of molecular-targeted drugs.Results:A total of1201 differentially expressed genes were identified,of which EGR1,MAPK1,MYC,BCL2 and CALM1 were important.The results of GO and KEGG analysis indicated that the formation of PRL was mainly related with changes of spindle pole and influenced oocyte meiosis.PRL was likely to have some homology with Parkinson’s disease.Simultaneously,CCK-8 assay,clone formation assay and in vitro wound-healing test confirmed that EGR1 agonist of Genipin had potential effect on anti-prolactinoma.Results of flow cytometry analysis showed that the apoptosis percentage of tumor cells was increased as the dose of Genipin increased.Conclusion:EGR1,MAPK1,MYC,BCL2 and CALM1 are core driving genes of PRL,which have important impacts on the PRL development.Genipin,an agonist of EGR1,inhibiting the proliferation and migration of PRL cells in vitro,shows a potential therapeutic effect on PRL.At the same time,this study screened a high-risk signaling pathway for the PRL development,providing a new target for clinical diagnosis and treatment,with great significance.