U2AF2表达对肝细胞癌增殖和迁移的影响及其与预后的关系  

Effect of U2AF2 expression on proliferation and migration of hepatocellular carcinoma and its relationship with prognosis

作　　者：莫钊鸿 翟航 苏日顺 孟泓宇 罗豪 陈文豪 许瑞云[1] Zhaohong;Zhai Hang;Su Rishun;Meng Hongyu;Luo Hao;Chen Wenhao;Xu Ruiyun(Department of Hepatobiliary Surgery,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院肝胆外科,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2023年第3期336-341,共6页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：广州市科技计划项目(201804010211)。

摘　　要：目的:探讨U2小核RNA辅助因子2(U2AF2)表达对肝细胞癌增殖和迁移的影响及其与预后的关系。方法:采用实时荧光定量PCR(qRT-PCR)检测U2AF2在HCC细胞系(HepG2、Hep3B、Huh-7和HCCLM9)及正常肝脏细胞LO2中的表达水平。采用慢病毒介导的短发夹RNA(shRNA)敲低U2AF2在HCC细胞中的表达(shU2AF2-1、shU2AF2-2为实验组,sh-NC为对照组)。采用CCK-8和细胞划痕实验分别检测shU2AF2-1组和sh-NC组HCC细胞的增殖和迁移能力。Western bolt检测shU2AF2-1组和sh-NC组HCC细胞的哺乳动物雷帕霉素靶蛋白(mTOR)表达水平及其磷酸化(p-mTOR)水平。生物信息学分析采用UALCAN和Kaplan-Meier Plotter数据库分别分析U2AF2在HCC中的表达及其与HCC预后的关系。下载癌症基因组图谱(TCGA)数据库的HCC表达谱,采用基因集富集分析(GSEA)分析U2AF2在HCC中参与的信号通路。多组比较采用单因素方差分析,两组比较采用LSD-t或t检验。结果:TCGA数据库分析显示,HCC组织U2AF2 mRNA表达水平明显高于癌旁正常肝组织(P<0.05),U2AF2 mRNA表达水平随着HCC肿瘤分级的增高而增高(P<0.05)。Kaplan-Meier Plotter数据库分析显示,U2AF2高表达组患者总体生存率及无复发生存率明显低于低表达组(HR=1.86,1.85;P<0.05)。GSEA结果提示,U2AF2的高表达与mTOR通路密切相关,标准化富集得分(NES)为2.113(P<0.05)。qRT-PCR检测显示,U2AF2 mRNA在HepG2、Hep3B、Huh-7和HCCLM9细胞中表达水平明显高于正常肝脏细胞LO2,其中以HCCLM9细胞的表达最高(F=26.003,P<0.05);shU2AF2-1和shU2AF2-2组HCCLM9细胞U2AF2 mRNA平均相对表达量分别为0.27±0.09、0.41±0.13,明显低于sh-NC组的1.02±0.06(LSD-t=-7.643,-6.220;P<0.05)。CCK-8实验显示,shU2AF2-1组HCCLM9细胞的增殖能力明显低于sh-NC组(t=-5.381,P<0.05)。细胞划痕实验显示,shU2AF2-1组HCCLM9细胞的迁移能力明显低于sh-NC组(t=-7.903,P<0.05)。Western blot检测显示,shU2AF2-1组HCCLM9细胞的p-mTOR水平明显低于sh-NC组,而两组mTOR水�Objective To evaluate the effect of U2 small nuclear RNA auxiliary factor 2(U2AF2)expression on the proliferation and migration of hepatocellular carcinoma and its relationship with prognosis.Methods The expression level of U2AF2 in HCC cell lines(HepG2,Hep3B,Huh-7 and HCCLM9)and normal liver cell LO2 was detected by quantitative real-time polymerase chain reaction(qRT-PCR).The short hairpin RNA(shRNA)mediated by lentivirus was employed to knock down the expression of U2AF2 in HCC cells(shU2AF2-1 and shU2AF2-2 were assigned into the experimental group and sh-NC as the control group).The proliferation and migration capability of HCC cells in the shU2AF2-1 and sh-NC groups were evaluated by CCK-8 assay and wound healing assay,respectively.The expression levels of mammalian target of rapamycin(mTOR)and phosphorylated mTOR(p-mTOR)in HCC cells in the shU2AF2-1 and sh-NC groups were measured by Western blot.Regarding bioinformatics analysis,the expression level of U2AF2 in HCC and its relationship with prognosis were analyzed by using UALCAN and Kaplan-Meier Plotter databases,respectively.The expression profile of HCC was downloaded from The Cancer Genome Atlas(TCGA)database.The signal pathways of U2AF2 involved in HCC were analyzed by Gene Set Enrichment Analysis(GSEA).Multi-group comparison was analyzed by one-way ANOVA.Two-group comparison was conducted by LSD-t or t test.Results TCGA database analysis showed that the expression level of U2AF2 mRNA in HCC tissues was significantly higher than that in adjacent normal liver tissues(P<0.05),and the expression level of U2AF2 mRNA was up-regulated with the increase of HCC grade(P<0.05).Kaplan-Meier plotter database analysis revealed that the overall survival and recurrence-free survival rates of patients with high expression of U2AF2 were significantly lower than those of their counterparts with low expression(HR=1.86,1.85;P<0.05).GSEA analysis indicated that high expression of U2AF2 was closely correlated with the mTOR signaling pathway,and the normalized enrichment sco

关 键 词：癌 肝细胞 U2小核RNA辅助因子2(U2AF2) 细胞增殖 细胞迁移 哺乳动物雷帕霉素靶蛋白(mTOR) 预后 

分 类 号：R735.7[医药卫生—肿瘤]