miR-129-5p通过靶向SALL4抑制肝癌细胞增殖、迁移和侵袭的实验研究  

The experimental study of miR-129-5p inhibiting proliferation,migration,and invasion of hepatocellular carcinoma by targeting to SALL4

作　　者：张华 范松 刘冀琴 张学军 ZHANG Hua;FAN Song;LIU Jiqin;ZHANG Xuejun(Department of Immunology,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China;Department of Clinical Laboratory,Characteristic Medical Center of Chinese People′s Armed Police Force,Tianjin 300162,China;Department of Clinical Laboratory,Armed Police Corps Hospital of Hebei Province,Shijiazhuang 050000,China)

机构地区：[1]天津医科大学基础医学院免疫学系,天津300070 [2]武警特色医学中心检验科,天津300162 [3]武警河北省总队医院检验科,石家庄050000

出　　处：《天津医科大学学报》2024年第1期11-14,34,共5页Journal of Tianjin Medical University

摘　　要：目的:探讨miR-129-5p在肝细胞癌(HCC)发展中的作用及其机制。方法:通过实时荧光定量-PCR(qRT-PCR)检测miR-129-5p在正常血清及肝癌患者血清、人正常肝细胞LO2和人肝癌细胞系中的表达情况。采用脂质体转染技术将miR-129-5p mimics转染至人肝癌细胞HCCLM3细胞,应用Western印迹检测人类婆罗双树样基因-4(SALL4)和程序性死亡受体-配体1(PD-L1)蛋白的表达。通过CCK-8实验、克隆形成实验、细胞周期检测、细胞划痕实验和Transwell实验等检测在体外过表达miR-129-5p对肝癌细胞增殖、迁移及侵袭能力的影响。此外,通过生物信息学工具、数据库分析和荧光素酶报告基因检测实验,探索miR-129-5p在HCC中的靶点,并探讨靶点SALL4是否介导miR-129-5p对HCC细胞的作用。结果:与正常血清相比,肝癌患者血清miRNA-129-5p表达量显著降低(t=13.32,P<0.001),与LO2相比,肝癌细胞系HepG2、Hep3B、HCCLM3、BEL-7402和QGY-7703的miR-129-5p表达量均显著降低(t=30.35、33.08、37.11、32.87、8.2,均P<0.05)。miR-129-5p过表达可以抑制肝癌细胞增殖、侵袭和转移。StarBase预测显示miR-129-5p与SALL4有潜在结合位点,双荧光素酶报告基因检测证明miR-129-5p与SALL4直接结合。与对照组相比,miR-129-5p表达后SALL4和PD-L1的蛋白水平明显降低(t=12.68、t=8.798,均P<0.05)。miR-129-5p可以通过调控SALL4的表达,抑制HCC细胞的活力、增殖、迁移和侵袭(F=26.11、147.2、4.302、321.3,均P<0.05)。结论:过表达miR-129-5p通过抑制SALL4表达,抑制肝癌细胞增殖、迁移和侵袭。Objective:To explore the role of miR-129-5p in hepatocellular carcinoma(HCC)development and its mechanism.Methods:The expression of miR-129-5p in normal serum and serum of hepatocellular carcinoma patients,human normal hepatocyte LO2 and human hepatocellular carcinoma cell line was detected by quantitative real-time PCR(qRT-PCR).Human hepatocellular carcinoma cells HCCLM3 were selected as the research objects,and miR-129-5p mimics was transfected into HCCLM3 cells by liposome transfection technology.Western blotting was used to detect the protein expression of Sal-like gene 4(SALL4)and programmed death receptor-ligand 1(PD-L1).CCK-8 assay,colony formation assay,cell cycle detection,cell scratch assay and Transwell assay were used to detect the effects of miR-129-5p overexpression on the proliferation,migration and invasion of HCC in vitro.In addition,bioinformatics tools,database analysis,luciferase reporter gene detection and rescue experiments were used to explore the target of miR-129-5p in HCC and to explore whether the target SALL4 mediated the effect of miR-129-5p on HCC cells.Results:The expression of miR-129-5p was signifi-cantly decreased in the serum of HCC patients compared with normal serum(t=13.32,P<0.001).Compared with LO2,the expression of miR-129-5p in liver cancer cell lines including HepG2,Hep3B,HCCLM3,BEL-7402 and QGY-7703 was significantly decreased(t=30.35,33.08,37.11,32.87,8.2,all P<0.05).Overexpression of miR-129-5p inhibited the proliferation,invasion and metastasis of hepatocellular carcinoma cells.StarBase predicted that miR-129-5p had potential binding sites with SALL4,and dual luciferase reporter gene assay confirmed that miR-129-5p directly bound to SALL4.Compared with the control group,the protein levels of SALL4 and PD-L1 were significantly decreased after miR-129-5p expression(t=12.68,8.798,both P<0.05).miR-129-5p could inhibit the viability,proliferation,migration and invasion of HCC cells by regulating the expression of SALL4(F=26.11,147.2,4.302,321.3,all P<0.05).Conclusion:Overe

关 键 词：肝细胞癌 miR-129-5p SALL4 增殖 侵袭 迁移 

分 类 号：R575.7[医药卫生—消化系统]