LncRNA FEZF1-AS1靶向miR-1254调控肝癌细胞的生物学行为  

LncRNA FEZF1-AS1 Regulates Biological Behavior of Hepatocellular Carcinoma Cells by Targeting MiR-1254

作　　者：田贵金[1] 刘军红 陈晶 张小龙 杨志森 TIAN Guijin;LIU Junhong;CHEN Jing;ZHANG Xiaolong;YANG Zhisen(Department of Surgery,the First Hospital of Handan,Handan,Hebei Province,056002;Department of Obstetrics,Wu’an First People’s Hospital,Wu’an,Hebei Province)

机构地区：[1]河北省邯郸市第一医院综合外科,056002 [2]河北省武安市第一人民医院产科

出　　处：《胃肠病学》2020年第5期288-294,共7页Chinese Journal of Gastroenterology

摘　　要：背景:肝癌是我国常见的恶性肿瘤之一,严重威胁人们生命健康。目前,越来越多研究证实长链非编码RNA(LncRNA)在癌症发生、进展中的作用。目的:探讨LncRNA FEZF1-AS1靶向miR-1254调控肝癌细胞生物学行为的分子机制。方法:采用qRT-PCR法检测肝癌组织和细胞株中FEZF1-AS1和miR-1254表达。将肝癌Hep3B细胞分为NC组、si-Lnc FEZF1-AS1组、si-con组、miR-1254组、miR-con组、si-Lnc FEZF1-AS1+anti-miR-con组、si-Lnc FEZF1-AS1+anti-miR-1254组。MTT法检测细胞活力,流式细胞术检测细胞凋亡,Transwell实验检测细胞迁移和侵袭能力,双萤光素酶报告基因实验验证FEZF1-AS1与miR-1254的靶向调控关系,蛋白质印迹法检测相关蛋白的表达。结果:肝癌组织和3种肝癌细胞株中FEZF1-AS1表达显著高于相应癌旁组织和正常细胞,miR-1254表达显著降低。转染si-Lnc FEZF1-AS1或miR-1254 mimics均可显著抑制Hep3B细胞Ki-67、N-cadherin和Vimentin表达,促进cleaved caspase-3和E-cadherin蛋白表达,抑制细胞增殖、迁移和侵袭能力,抑制EMT发生,促进细胞凋亡。FEZF1-AS1靶向负调控miR-1254表达。转染si-Lnc FEZF1-AS1可抑制PI3K/AKT信号通路活化。转染anti-miR-1254可逆转转染si-Lnc FEZF1-AS1对Hep3B细胞增殖、迁移、侵袭、EMT、凋亡以及PI3K/AKT信号通路的影响。结论:LncRNA FEZF1-AS1在肝癌患者中的表达上调。沉默LncRNA FEZF1-AS1通过靶向miR-1254抑制肝癌细胞增殖、迁移、侵袭以及EMT发生,促进细胞凋亡,其机制与抑制PI3K/AKT信号通路活化有关。Background:Hepatocellular carcinoma is one of the most common malignant tumors in China,which seriously threatens the life of patients.Nowadays,more and more studies have confirmed the role of long non-coding RNA(LncRNA)in the development and progression of cancer.Aims:To investigate the molecular mechanism of FEZF1-AS1 regulates the biological behavior of hepatocellular carcinoma cells by targeting miR-1254.Methods:qRT-PCR was used to detect the expressions of FEZF1-AS1 and miR-1254 in hepatocellular carcinoma tissue and cells.Hepatocellular carcinoma Hep3B cells were randomly divided into NC group,si-Lnc FEZF1-AS1 group,si-con group,miR-1254 group,miR-con group,si-Lnc FEZF1-AS1+anti-miR-con group,si-Lnc FEZF1-AS1+anti-miR-1254 group.The cell viability was detected by MTT assay,apoptosis was detected by flow cytometry,the cell migration and invasion ability were detected by Transwell assay,the dual luciferase reporter gene assay was used to verify the targeted regulatory relationship between FEZF1-AS1 and miR-1254,Western blotting was used to detect the expressions of related proteins.Results:The expression of FEZF1-AS1 was significantly increased whereas the expression of miR-1254 was significantly decreased in hepatocellular carcinoma tissue and cells than those in corresponding adjacent tissue and normal cells.Transfection of si-Lnc FEZF1-AS1 or miR-1254 mimics significantly inhibited the expressions of Ki-67,N-cadherin and Vimentin in Hep3B cells,promoted the expressions of cleaved caspase-3 and E-cadherin,inhibited cell proliferation,migration and invasion ability,inhibited the occurrence of EMT and promoted cell apoptosis.FEZF1-AS1 negatively regulated miR-1254 expression.Transfection of si-Lnc FEZF1-AS1 inhibited activation of PI3K/AKT signaling pathway.Transfection of anti-miR-1254 reversed the effect of si-Lnc FEZF1-AS1 transfection on Hep3B cell proliferation,migration,invasion,EMT,apoptosis and PI3K/AKT signaling pathway.Conclusions:The expression of LncRNA FEZF1-AS1 is up-regulated in patients with h

关 键 词：FEZF1-AS1 MiR-1254 肝肿瘤 细胞增殖 细胞迁移 细胞侵袭 细胞凋亡 上皮-间质转化 

分 类 号：R73[医药卫生—肿瘤]