敲低CCAAT/增强子结合蛋白β促进肝癌细胞的增殖和迁移  

Knockdown of C/EBPβPromotes the Proliferation and Migration of Hepatocellular Carcinoma Cells

作　　者：陈利荣 刘玉玲 贾艳梅 梁紫微 李佳佳 CHEN Li-Rong;LIU Yu-Ling;JIA Yan-Mei;LIANG Zi-Wei;LI Jia-Jia(Department of Medical Laboratory,Fenyang College,Shanxi Medical University,Fenyang 032200,Shanxi,China;School of Biomedical Engineering,Taiyuan University of Technology,Taiyuan 030000,China;Graduate School,Shanxi Medical University,Taiyuan 030000,China)

机构地区：[1]山西医科大学汾阳学院医学检验系,山西汾阳032200 [2]太原理工大学生物医学工程学院,太原030000 [3]山西医科大学研究生学院,太原030000

出　　处：《中国生物化学与分子生物学报》2024年第6期797-806,共10页Chinese Journal of Biochemistry and Molecular Biology

基　　金：山西省吕梁市科学技术局社会发展领域重点研发计划项目(No.2020SHFZ34);国家自然科学基金项目(No.82103147)资助。

摘　　要：近年来已有研究表明,C/EBPβ在肝癌的发生发展中发挥重要的作用,但其具体分子调控机制尚不清楚。本研究通过分析GEO数据库和Kaplan-Meier Plotter数据库发现,C/EBPβmRNA在肝癌中低表达(P<0.05),且其低表达与肝癌患者预后密切相关。进一步,通过功能富集分析和TIMER数据库分析显示,C/EBPβ主要参与细胞周期、DNA转录等生物学过程,且C/EBPβ表达水平与CD4^(+)T细胞和巨噬细胞的免疫浸润具有较强的相关性(P<0.05)。为了进一步研究C/EBPβ对肝癌细胞增殖和迁移的影响。在肝癌细胞中瞬时转染C/EBPβsiRNA,将其分为si-NC组和siC/EBPβ组。通过qRT-PCR、Western印迹检测肝癌细胞中C/EBPβ在mRNA和蛋白质水平均显著降低(P<0.05);通过MTT检测、平板克隆形成实验和5-乙炔基-2-脱氧尿嘧啶核苷(Edu)实验证实,敲低C/EBPβ促进肝癌细胞的增殖(P<0.05);Transwell和划痕实验证实,敲低C/EBPβ促进肝癌细胞的迁移;Western印迹法检测敲低C/EBPβ对肝癌细胞内迁移相关蛋白质(E-cadherin、N-cadherin)以及Wnt/β-catenin信号通路蛋白质表达的影响,结果表明,敲低C/EBPβ促进肝癌细胞EMT上皮间质转化,并能激活Wnt/β-catenin信号通路的基因表达(P<0.05)。综上所述,C/EBPβ在肝癌组织中低表达且与患者生存预后成正相关。敲低C/EBPβ可能通过激活Wnt/β-catenin信号通络促进肝癌细胞的增殖、迁移和上皮间质转化,为C/EBPβ在肝癌的发生发展中的作用提供了依据,可能是一个肝癌诊治过程中的潜在靶点。In recent years,studies have shown that C/EBPβplays an important role in the occurrence and development of liver cancer,but its specific molecular regulatory mechanism is still unclear.In this study,we analyzed the GEO database and the Kaplan-Meier Plotter database and found that the mRNA expression of C/EBPβwas low in hepatocellular carcinoma(HCC)cells(P<0.05),and its low expression was closely related to the prognosis of HCC patients.Furthermore,functional enrichment analysis and TIMER database analysis showed that C/EBPβwas mainly involved in biological processes such as cell cycle and DNA transcription,and the expression level of C/EBPβhad a strong correlation with the immune infiltration of CD4^(+)T cells and macrophages(P<0.05).To further investigate the effect of C/EBPβon the proliferation and migration of HCC cells.In the experiment,HCC cells were transiently transfected with C/EBPβsiRNA and divided into si-NC group and siC/EBPβgroup.The mRNA and protein levels of C/EBPβin HCC cells were significantly reduced by qRT-PCR and Western blot(P<0.05),and MTT detection,plate cloning assay,and 5-ethynyl-2’deoxyuracil nucleoside(Edu)assay confirmed that knockdown of C/EBPβpromotes the proliferation of HCC cells(P<0.05);Transwell and scratch assays confirmed that knockdown of C/EBPβpromote the migration of HCC cells;Western blot method was used to detect the effect of knockdown of C/EBPβon the expression of migration-related proteins(E-cadherin,N-cadherin)and Wnt/β-catenin signaling pathway proteins.The results showed that knockdown of C/EBPβpromotes epithelial-mesenchymal transition(EMT)and activates gene expression of Wnt/β-catenin signaling pathway in HCC cells(P<0.05).In conclusion,C/EBPβwas underexpressed in HCC tissues and was positively correlated with the survival prognosis of patients.Knockdown of C/EBPβmay promote the proliferation,migration,and epithelial-mesenchymal transformation of HCC cells by activating the Wnt/β-catenin signaling pathway,which provides a basis for the role of C/E

关 键 词：肝癌 细胞增殖 细胞迁移 CCAAT/增强子结合蛋白β 

分 类 号：Q71[生物学—分子生物学]