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/EBP β Promotes the Proliferation and Migration of Hepatocellular Carcinoma Cells
陈利荣 1刘玉玲 1贾艳梅 1梁紫微 2李佳佳3
作者信息
- 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. 山西医科大学汾阳学院医学检验系,山西汾阳 032200;山西医科大学研究生学院,太原 030000
- 折叠
摘要
近年来已有研究表明,C/EBP β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/EBP β mRNA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/EBP β主要参与细胞周期、DNA转录等生物学过程,且C/EBP β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/EBP β siRNA,将其分为si-NC组和siC/EBP β组.通过qRT-PCR、Western印迹检测肝癌细胞中C/EBP β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/EBP β促进肝癌细胞的增殖(P＜0.05);Transwell和划痕实验证实,敲低C/EBP β促进肝癌细胞的迁移;Western印迹法检测敲低C/EBP β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/EBP β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/EBP β在肝癌组织中低表达且与患者生存预后成正相关.敲低C/EBP β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/EBP β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.
Abstract
In recent years,studies have shown that C/EBP β plays 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/EBP β was low in hepatocellular carcinoma(HCC)cells(P＜0.05),and its low ex-pression was closely related to the prognosis of HCC patients.Furthermore,functional enrichment analy-sis and TIMER database analysis showed that C/EBP β was mainly involved in biological processes such as cell cycle and DNA transcription,and the expression level of C/EBP β had a strong correlation with the immune infiltration of CD4+T cells and macrophages(P＜0.05).To further investigate the effect of C/EBP β on the proliferation and migration of HCC cells.In the experiment,HCC cells were transiently transfected with C/EBP β siRNA and divided into si-NC group and siC/EBP β group.The mRNA and protein levels of C/EBP β in 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/EBP β promotes the proliferation of HCC cells(P＜0.05);Transwell and scratch assays confirmed that knockdown of C/EBP β promote the migration of HCC cells;Western blot method was used to detect the effect of knockdown of C/EBP β on the expression of migration-related proteins(E-cadherin,N-cadherin)and Wnt/β-catenin signaling pathway proteins.The results showed that knockdown of C/EBP β promotes epithelial-mesenchymal transition(EMT)and activates gene ex-pression of Wnt/β-catenin signaling pathway in HCC cells(P＜0.05).In conclusion,C/EBP β was un-derexpressed in HCC tissues and was positively correlated with the survival prognosis of patients.Knock-down of C/EBP β may 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/EBP β in the occurrence and development of HCC and may be a potential target in the diagnosis and treatment of HCC.
关键词
肝癌/细胞增殖/细胞迁移/CCAAT/增强子结合蛋白βKey words
hepatocellular carcinoma(HCC)/cell proliferation/cell migration/CCAAT/enhancer binding protein β(C/EBP β)引用本文复制引用
基金项目
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(2020SHFZ34)
国家自然科学基金项目(82103147)
出版年
2024
NETL
NSTL
万方数据