装载miRNA-451a的外泌体对肝癌细胞株增殖的影响研究  

Effect of exosomes loaded with miRNA-451a on the proliferation of hepatocellular carcinoma cell lines in vitro

作　　者：王壮 朱冬梅[1] 申红玉 叶春艳 WANG Zhuang;ZHU Dong-mei;SHEN Hong-yu(Changzhou School of Clinical Medicine,Nanjing Medical University,Changzhou,Jiangshu 213001)

机构地区：[1]南京医科大学常州医学中心,常州市第三人民医院肝病科,常州213001

出　　处：《肝胆外科杂志》2023年第4期309-313,共5页Journal of Hepatobiliary Surgery

基　　金：常州市科学技术局应用基础研究计划项目(编号:CJ20179055、CJ20190077、CJ20210088)。

摘　　要：目的探讨装载miRNA451a的外泌体对肝癌细胞株HepG2和SMMC-7721增殖的影响。方法正常肝组织上皮细胞株HL-7702来源的外泌体,通过与胆固醇基修饰的miRNA-451a模拟物直接孵育构建外泌体451a复合体。将构建的exo-miRNA-451a复合体、等量miRNA-451a模拟物、等量PBS分别接种于96孔板中处于对数生长期的HPG2和SMMC-7721细胞,作为实验组、对照组和空白对照组。采用激光共聚焦显微镜观察exo-miRNA-451a复合体进入细胞内的情况;CCK8法检测HPG2和SMMC-7721细胞增殖。结果构建的外泌体miRNA-451a复合体分别与HepG2和SMMC-7721细胞,共培养3h时,在共聚焦显微镜下,有PKH67标记的外泌体和Cy3标记的miRNA-451a在HepG2和SMMC-7721细胞中共定位,并且在随后的6h、12h观察时逐渐增多。CCK-8实验提示当外泌体451a复合体和miR-451a模拟物分别与肝癌细胞株共孵育6h,OD值差异不明显;共孵育12h实验组OD值低于对照组,共培养48h实验组OD值显著低于对照组。结论装载miRNA-451a的外泌体可抑制肝癌细胞株的增殖。Objective To explore the effect of exosomes loaded with miRNA451a on the proliferation of hepatocellular carcinoma cell lines HepG2 and SMMC-7721.Methods Exosomes derived from normal liver tissue epithelial cell line HL-7702 were used to construct the exosome 451a complex by direct incubation with cholesterol-based modified miRNA-451a mimics.The exo-miRNA-451a complex,equal amounts of miRNA-451a mimics,and equal amounts of PBS were inoculated into HPG2 and SMMC-7721 cells in logarithmic growth phase in 96-well plates as experimental,control and blank control groups.Laser confocal microscopy was used to survey the entry of exo-miRNA-451a complex into the cells;the proliferation of HepG2 and SMMC-7721 cells was detected by CCK8 method.Results The constructed exo-miRNA-451a complex was co-cultured with HepG2 and SMMC-7721 cells,respectively,for 3 hours,with PKH67-labeled exosome and Cy3-labeled miRNA-451a co-localized in HepG2 and SMMC-7721 cells under confocal microscopy,and gradually increased when observed at the subsequent 6 hours and 12 hours.CCK-8 experiments suggested that when the exosome 451a complex and miR-451a simulants were co-incubated with hepatoma cell lines for 6 hours,respectively,the difference in OD was not significant;the OD of experimental group was lower than control group when co-incubated for 12 h,the OD of experimental group was significantly lower than control group when co-incubated for 48 h.Conclusion The exosomes loaded with miRNA-451a could inhibit the proliferation of hepatoma cells.

关 键 词：外泌体 微小RNA-451a 肝癌细胞株 

分 类 号：R735.7[医药卫生—肿瘤]