摘要
目的 筛选苹果酸酶2(ME2)调控的基因并探究其在肝癌发生发展中的作用与机制。方法 对ME2敲降细胞的RNA-seq数据进行差异基因分析、聚类分析、GO和KEGG富集分析;在HepG2细胞中通过荧光定量PCR检测ME2敲降或者过表达后候选靶基因SHCBP1 mRNA的水平;Western blot检测ME2对下游信号通路的影响;通过细胞增殖、划痕与集落形成实验检测SHCBP1对肝癌细胞增殖与迁移的影响;利用TCGA LIHC数据库中肝癌患者的生存数据分析SHCBP1对肝癌患者预后的影响。结果 敲降ME2后差异基因富集于PI3K-Akt信号通路、细胞周期、丝氨酸磷酸化等生物过程。在HepG2中敲降ME2后SHCBP1的mRNA水平降低,过表达ME2后SHCBP1的mRNA水平升高,两者呈正相关。Western blot结果显示ME2通过SHCBP1激活PI3K-Akt信号通路。荧光定量PCR结果显示SHCBP1在肝癌细胞系中表达显著高于正常肝细胞,并促进肝癌细胞的增殖与迁移,且与不良预后相关。结论 ME2正向调控SHCBP1并激活PI3K-Akt信号通路,SHCBP1高表达促进肝癌的发生发展,为肝癌治疗提供了新的靶点。
Objective To identify genes regulated by ME2 and to explore their roles as well as underlying mechanisms in liver cancer progression.Methods RNA-seq data of siME2-transfected cells were subjected to differential expression analysis,clustering,GO and KEGG enrichment analyses.The mRNA level of the potential target gene SHCBP1 was measured by quantitative real-time PCR(qPCR)following ME2 knockdown or overexpression in HepG2 cells.The effect of ME2 and SHCBP1 on the downstream pathway was examined by Western blot.Cell proliferation,wound healing,and colony formation assays were conducted to evaluate SHCBP1′s role in liver cancer cell proliferation and migration.Survival analysis of the TCGA-LIHC cohort was performed to determine the prognostic value of SHCBP1 in liver cancer patients.Results Differentially expressed genes in siME2-transfected cells were significantly enriched in biological processes including the PI3K-Akt signaling pathway,cell cycle,and serine phosphorylation.In HepG2 cells,ME2 knockdown led to a reduction in SHCBP1 mRNA level,whereas ME2 over-expression resulted in enhanced SHCBP1 mRNA level,demonstrating a positive correlation between ME2 and SHCBP1 expression.Western blot analysis revealed that ME2 enhanced PI3K-Akt signaling pathway activation through SHCBP1.qPCR results confirmed that SHCBP1 was significantly over-expressed in liver cancer cells and promoted both proliferation and migration,contributing to poor prognosis in liver cancer patients.Conclusions ME2 promotes liver cancer progression by regulating SHCBP1 to activate the PI3K-Akt signaling pathway,presenting a novel therapeutic target for liver cancer treatment.
作者
秋昱翀
杜文静
QIU Yuchong;DU Wenjing(State Key Laboratory of Common Mechanism Research for Major Diseases,Department of Cell Biology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005;4+4 Medical Doctor Program,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100730,China)
出处
《基础医学与临床》
2025年第6期741-747,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(32470829)
中国医学科学院医学与健康科技创新工程(2021-I2M-1-016)。
