miR-424-5p靶向KIF23调控细胞周期并抑制肝癌细胞的增殖和迁移能力

MiR-424-5p targets KIF23 to modulate the cell cycle and impede the proliferation and migration of hepatocellular carcinoma cells

目的探究miR-424-5p、KIF23对肝癌细胞的作用机制,为探索新的肝癌靶向治疗途径提供思路。方法实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-424-5p的表达水平,生物信息学技术和萤光素酶报告基因验证KIF23与miR-424-5p在肝癌中的结合关系。qRT-PCR和蛋白免疫印迹(western blot,WB)分别检测KIF23 mRNA和蛋白表达水平。CCK8实验、克隆形成实验、迁移实验和流式细胞术评估miR-424-5p和KIF23对肿瘤细胞的增殖、迁移能力,以及对细胞周期进展的影响。结果与正常肝组织比较,miR-424-5p在肝癌组织和细胞中显著低表达(P<0.05);KIF23是miR-424-5p的直接下游靶点,在肝癌组织和细胞中的表达显著上调(P<0.05)。过表达KIF23会促进肝癌细胞增殖和迁移,并将细胞周期阻滞于G2/M期。过表达miR-424-5p能显著减弱过表达KIF23对细胞增殖、迁移和细胞周期分布的影响(P<0.05)。结论miR-424-5p可以通过靶向下调KIF23,从而抑制肝癌细胞增殖和迁移能力,影响细胞周期进展。

Objective The aim of this study is to investigate the mechanistic role of miR-424-5p and KIF23 in Hepatocellular Carcinoma cells,with a view to providing new perspectives on targeted therapeutic strategies for liver cancer.Methods qRT-PCR was employed to quantify the mRNA expression level of miR-424-5p,while bioinformatics technology and luciferase reporter gene assay were utilized to validate the binding relationship between KIF23 and miR-424-5p in Hepatocellular Carcinoma cells.qRT-PCR and western blot analyses were employed to assess the mRNA and protein expression levels of KIF23.The effects of miR-424-5p and KIF23 on cell proliferation,migration ability,and cell cycle progression were evaluated using CCK8 assay,clone formation assay,migration assay,and flow cytometry.Results Compared to normal liver tissues,the expression of miR-424-5p was significantly downregulated in Hepatocellular Carcinoma tissues and cells(P<0.05).KIF23 was identified as a direct downstream target of miR-424-5p,and its expression level was significantly upregulated in Hepatocellular Carcinoma tissues and cells(P<0.05).The overexpression of KIF23 promotes the proliferation and migration of Hepatocellular Carcinoma cells while inducing cell cycle arrest at the G2/M phase.Meanwhile,the overexpression of miR-424-5p significantly attenuated the impact of KIF23 on cellular proliferation,migration,and cell cycle distribution(P<0.05).Conclusions miR-424-5p was found to down-regulate KIF23,thereby inhibiting the proliferation and migration abilities of Hepatocellular Carcinoma cells while also affecting cell cycle progression.