缺氧抑制因子ZNF92在肝癌生长和侵袭中的功能研究  

Functional Study of Hypoxia-inhibitory Factor ZNF92 in the Growth and Invasion of Hepatocellular Carcinoma

作　　者：李婷婷 张婷 张亚楠 刘婕 丁丽华 叶棋浓 LI Tingting;ZHANG Ting;ZHANG Yanan;LIU Jie;DING Lihua;YE Qinong(School of Medicine,Guizhou University,Guiyang 550000,China;Institute of Bioengineering,Institute of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]贵州大学医学院,贵阳550000 [2]军事科学院军事医学研究院生物工程研究所,北京100850

出　　处：《中国生物工程杂志》2024年第11期20-29,共10页China Biotechnology

摘　　要：目的:探究缺氧调控因子锌指蛋白92(zinc finger proteins 92,ZNF92)在肝细胞癌生长和侵袭中发挥的功能及其可能的调控机制。方法:(1)利用RNA-seq筛选出缺氧抑制因子ZNF92并通过RT-qPCR和Western blot检测缺氧环境(1%O2)对ZNF92的mRNA表达水平和蛋白质表达水平的影响。(2)利用慢病毒载体构建敲低ZNF92基因的肝癌细胞系。(3)通过CCK-8实验、划痕试验和Transwell实验确定ZNF92基因对肝癌细胞HepG2、SMMC7721生长和侵袭的影响。(4)通过在敲低ZNF92基因的稳定肝癌细胞系中瞬时转染锌指E盒结合同源盒1 (zinc finger E-box binding homeobox, ZEB1)的siRNA,通过Western blot确定ZEB1转染成功,并在该细胞系中检测ZNF92能否通过调控ZEB1的表达调控上皮间充质转化(epithelial-mesenchymal transition, EMT)进程。(5)通过在敲低ZNF92基因的稳定肝癌细胞系中瞬时转染缺氧诱导因子1α(hypoxia-inducible factor 1α,HIF-1α)的siRNA,检测ZNF92是否可能通过HIF-1α影响细胞迁移和侵袭。结果:(1)缺氧抑制ZNF92的mRNA表达和蛋白质表达。(2)ZNF92调控EMT的关键转录因子ZEB1的蛋白质表达影响EMT进程。(3)ZNF92抑制肝癌细胞迁移和侵袭,但不影响肝癌细胞增殖。(4)ZNF92通过ZEB1部分调控肝癌细胞迁移和侵袭。(5)ZNF92可能通过HIF-1α调控ZEB1以影响肝癌细胞迁移和侵袭。结论:新缺氧抑制因子ZNF92可能通过HIF-1α影响EMT进程,进而通过调控ZEB1影响肝癌细胞的迁移和侵袭。Objective:To investigate the function and mechanism of the hypoxia-regulated factor zinc finger proteins 92(ZNF92)in hepatocellular carcinoma cell growth and invasion.Methods:(1)The mRNA and protein expression levels of ZNF92 were detected by RT-qPCR and Western blot.(2)ZNF92 knockdown hepatocellular carcinoma cell line was established using lentiviral vectors.(3)The effects of ZNF92 on hepatocellular carcinoma HepG2 cells,and SMMC7721 cell growth and invasion in vitro were evaluated by CCK-8,scratch and transwell assays.(4)Small interfering RNA(siRNA)was used to determine the knockdown of ZEB1 expression and to determine whether ZNF92 indirectly regulates the epithelial-mesenchymal transition(EMT)process through ZEB1.(5)siRNA was used to determine the knockdown of HIF-1αand to detect the changes in cell migration and invasive ability.Results:(1)Hypoxia inhibited the mRNA and protein expression of ZNF92.(2)ZNF92 regulates the protein expression of zinc finger E-box binding homeobox 1(ZEB1),a key transcription factor of EMT,thereby affecting the EMT process.(3)ZNF92 inhibits hepatocellular carcinoma migration and invasion without affecting the proliferation process.(4)ZNF92 partially regulates migration and invasion of hepatocellular carcinoma cells through ZEB1.(5)ZNF92 may partially regulate ZEB1 through HIF-1αto affect the migration and invasion of hepatocellular carcinoma cells.Conclusions:The novel hypoxia-inhibitory factor ZNF92 may affect the EMT process through HIF-1α,and then affect the migration and invasion of hepatocellular carcinoma cells by regulating ZEB1.

关 键 词：肿瘤缺氧 锌指蛋白92 缺氧诱导因子1Α 锌指E盒结合同源盒1 

分 类 号：Q816[生物学—生物工程]