miR-491-5p靶向FGFR4调节肝母细胞瘤细胞增殖和迁移

目的 探讨miR-491-5p靶向调节FGFR4对肝母细胞瘤增殖和迁移的影响及可能的作用机制。方法 采用qRT-PCR检测15例小儿肝母细胞瘤组织及其癌旁正常组织，正常肝细胞LO2和肝母细胞瘤细胞HuH-6中miR-491-5p和FGFR4 mRNA的表达；Western blot检测FGFR4蛋白表达；TargetScan数据库预测miR-491-5p靶点；双荧光素酶报告基因实验检测miR-491-5p与FGFR4的靶向关系。将mimic NC、miR-491-5p mimic、pcDNA3.1和pcDNA3.1-FGFR4质粒分别转染至HuH-6细胞后，采用qRT-PCR与Western blot检测FGFR4的表达，CCK-8实验检测细胞增殖能力，Transwell小室实验检测细胞迁移能力，Western blot检测GSK3β/β-catenin信号通路相关蛋白的表达。结果 小儿肝母细胞瘤组织和HuH-6细胞中miR-491-5p表达水平均低于癌旁正常组织及肝细胞LO2（P<0.05），而FGFR4表达水平高于癌旁正常组织及肝细胞LO2（P<0.05）；TargetScan数据库预测显示miR-491-5p与FGFR4存在结合位点。与mimic NC组比较，过表达miR-491-5p可抑制FGFR4表达、HuH-6细胞增殖和迁移能力及p-GSK3β/GSK3β、p-β-catenin/β-catenin及p-C-myc/C-myc比值（均P<0.05）。与mimic+Pc组比较，过表达FGFR4可逆转过表达miR-491-5p对HuH-6细胞增殖、迁移及p-GSK3β/GSK3β、p-β-catenin/β-catenin和p-C-myc/C-myc比值的抑制作用（均P<0.05）。结论 miR-491-5p通过负向调控FGFR4抑制肝母细胞瘤细胞增殖和迁移，可能与GSK3β/β-catenin信号通路失活有关。 

miR-491-59 regulates the proliferation and migration of hepatoblastoma cells by targeting FGFR4

Objective To investigate the effect of miR-491-5p targeting and regulating FGFR4 on the proliferation and migration of hepatoblastoma and its possible mechanism.Methods The expressions of miR-491-5p and FGFR4 m RNA in 15 cases of pediatric hepatoblastoma tissues and adjacent normal tissues,the normal hepatocytes cells LO2 and hepatoblastoma cells Hu H-6 were detected by q RT-PCR,and the expression level of FGFR4 protein was detected by Western blot.TargetScan database predicted the target of miR-491-5p,and the relationship between miR-491-5 and FGFR4 was detected by luciferase activity assay.After the mimic NC,miR-491-5p mimic,pcDNA3.1 plasmid and pcDNA3.1-FGFR4 plasmid were all transfected into Hu H-6 cells,the expression of FGFR4 was detected by q RT-PCR and Western blot,the cells proliferation was detected by CCK-8,the cell migration assay was detected by Transwell assay,and the expression of GSK3β/β-catenin signaling pathway related proteins was detected by Western blot.Results The expression level of miR-491-5p in pediatric hepatoblastoma tissues and Hu H-6 cells was lower than that in adjacent normal tissues and hepatocyte cells LO2(P<0.05),while the expression level of FGFR4 was higher than that in adjacent normal tissues and hepatocyte cells LO2(P<0.05).TargetScan database predicted that miR-491-5p and FGFR4 had binding sites.Compared with the mimic NC group,overexpression of miR-491-5p inhibited the expression of FGFR4,the proliferation and migration of hepatoblastoma cells HuH-6,and the ratios of p-GSK3β/GSK3β,p-β-catenin/β-catenin and p-C-myc/C-myc(all P<0.05).Compared with mimic+Pc group,overexpression of FGFR4 could reverse the inhibitory effect of miR-491-5p on the proliferation and migration of Hu H-6 cells,and the ratio of p-GSK3β/GSK3β,p-β-catenin/β-catenin and p-C-myc/C-myc(all P<0.05).Conclusion miR-491-5p negatively regulates FGFR4 to inhibit the proliferation and migration of hepatoblastoma cells,which may be related to the inactivation of GSK3β/β-catenin signaling pathway.