LncRNA-HULC靶向miR-597/Rab23分子轴影响肝癌细胞生物学特性的机制研究

Mechanism of LncRNA-HULC in Affecting the Biological Characteristics of Liver Cancer Cells by Targeting the miR-597/Rab23 Molecular Axis

该文旨在探讨长链非编码RNA(LncRNA)-肝癌高表达基因(HULC)靶向微小RNA-597(mi R-597)/Ras相关蛋白23(Rab23)分子轴影响肝细胞癌(HCC)细胞生物学特性的机制。以湖北医药学院附属襄阳市第一人民医院就诊的罹患HCC的76例患者为研究对象,qRT-PCR检测HCC患者组织中Lnc RNA-HULC、mi R-597及Rab23表达水平;并分析上述基因的表达水平与HCC的临床相关性;生物信息学方法和双荧光素酶报告实验分析Lnc RNA-HULC、mi R-597、Rab23信号轴的相互作用机制。以HCC细胞Hep G2为研究对象,将其分为si-NC组、si-HULC组、si-HULC+anti-NC组、siHULC+anti-mi R-597组,分别采用CCK8、Transwell实验、流式细胞仪检测各组Hep G2细胞的增殖、迁移、侵袭和凋亡情况;采用Western blot检测Hep G2细胞中Ki67、Caspase-3、Caspase-9、Rab23蛋白的表达情况。结果发现,HCC组织中Lnc RNA-HULC、Rab23的表达水平高于癌旁组织,miR-597的表达水平低于癌旁组织(P<0.05),且其表达情况与患者的TNM分期、分化程度等临床病理特征相关(P<0.05)。Lnc RNA-HULC靶向负调控mi R-597,miR-597靶向负调控Rab23。si-HULC组Lnc RNAHULC、Rab23 mRNA、增殖率、迁移数、侵袭数、Ki67蛋白、Rab23蛋白的表达水平低于si-NC组,mi R-597、凋亡率、Caspase-3蛋白、Caspase-9蛋白的表达水平高于si-NC组(P<0.05);与si-HULC组、si-HULC+anti-NC组比较,si-HULC+anti-miR-597组中mi R-597、凋亡率、Caspase-3蛋白、Caspase-9蛋白表达水平降低(P<0.05),Rab23 mRNA、增殖率、迁移数、侵袭数、Ki67蛋白、Rab23蛋白表达水平升高(P<0.05)。沉默Lnc RNA-HULC可能抑制肝癌细胞生物学特性,其机制可能是通过靶向mi R-597/Rab23分子轴实现的。

The aim of this study is to investigate the mechanism by which LncRNA(long non-coding RNA)-HULC(highly up-regulated in liver cancer)affects the biological characteristics of HCC(hepatocellular carcinoma)cells by targeting miR-597(microRNA-597)/Rab23(Ras related protein 23)molecular axis.A total of 76 patients with HCC who were admitted to the First People’s Hospital of Xiangyang,Hubei University of Medicine as the study subjects.QRT-PCR was applied to detect the expression levels of LncRNA-HULC,miR-597,and Rab23 in HCC patient tissues,and the expression of the aforementioned genes was analyzed for their clinical correlation with liver cancer.Bioinformatics methods and dual luciferase reporter experiments were applied to analyze the interaction mechanisms of LncRNA-HULC,miR-597,and Rab23 signaling axes.HepG2 HCC cells were studied and separated into si-NC group,si-HULC group,si-HULC+anti-NC group,and si-HULC+anti-miR-597 group.CCK8,Transwell assay,and flow cytometry were applied to detect the proliferation,migration,invasion,and apoptosis of HepG2 cells in each group.Western blot was applied to detect the expression of Ki67,Caspase-3,Caspase-9,and Rab23 proteins in HepG2 cells.The expression of LncRNA-HULC and Rab23 in HCC tissue was higher than that in adjacent tissues,while the expression of miR-597 was lower than that in adjacent tissues(P<0.05),and their expression was correlated with clinical and pathological features such as TNM stage and differentiation degree of patients(P<0.05).LncRNA-HULC targeted and negatively regulated miR-597,while miR-597 targeted and negatively regulated Rab23.The LncRNA-HULC,Rab23 mRNA,proliferation rate,migration number,invasion number,and the expression levels of Ki67 protein and Rab23 protein in the si-HULC group were lower than those in the si-NC group,and the miR-597,apoptosis rate,and the expression levels of Caspase-3 protein and Caspase-9 protein were higher than those in the si-NC group(P<0.05).Compared with the si-HULC group and si-HULC+anti-NC group,the miR-597 mRNA,apoptosis rate,and the expression levels of Caspase-3 protein and Caspase-9 protein in the si-HULC+anti-miR-597 group were lower(P<0.05),the Rab23 mRNA,proliferation rate,migration number,invasion number,and the expression levels of Ki67 protein and Rab23 protein were higher(P<0.05).In summary,silencing LncRNA-HULC may inhibit the biology of liver cancer cells,which may be achieved by targeting the miR-597/Rab23 molecular axis.