实用肿瘤杂志

目的 探究环状RNA circFBLIM1对肝癌细胞增殖、凋亡和迁移的调控作用，阐明其通过miR-338/磷酸果糖激酶肝型（phosphofructokinase liver type, PFKL）介导糖酵解影响肝癌进展的分子机制，为肝癌代谢靶向干预提供新的理论依据。方法 采用定量聚合酶链反应（quantitative polymerase chain reaction, qPCR）检测circFBLIM1在人肝癌细胞SNU-387与Huh-7及人正常肝细胞THLE-2中的表达水平。在SNU-387细胞中利用siRNA沉默或利用pcDNA载体过表达circFBLIM1。通过流式细胞术、细胞增殖-毒性检测试剂盒（Cell Counting Kit-8, CCK-8）和细胞划痕实验分析细胞凋亡、增殖和迁移能力。采用qPCR和Western blot分析circFBLIM1沉默和过表达对细胞中miR-338和PFKL表达的影响。结果 circFBLIM1在肝癌细胞SNU-387与Huh-7中高表达（均P<0.01）。在SNU-387细胞中沉默circFBLIM1可抑制细胞增殖，促进细胞凋亡，减弱细胞迁移能力，并上调miR-338表达，下调PFKL的m RNA和蛋白表达（均P<0.05）；过表达circFBLIM1则促进细胞增殖，增强迁移能力，下调miR-338表达，上调PFKL的mRNA和蛋白表达（均P<0.05）。结论 肝癌细胞中circFBLIM1高表达。circFBLIM1促进肝癌细胞增殖和迁移，并通过激活糖酵解通路circFBLIM1/miR-338/PFKL轴在肝癌进展中发挥作用。

Objective To investigate the regulatory effect of circular RNA circFBLIM1 on the proliferation, apoptosis and migration of liver cancer cells, and elucidate the molecular mechanism by which it affects liver cancer progression via miR-338/phosphofructokinase liver type(PFKL)-mediated glycolysis, so as to provide a novel theoretical basis for targeted metabolic intervention of liver cancer. Methods Quantitative polymerase chain reaction(qPCR) was used to detect the expression of circFBLIM1 in two human liver cancer cell lines(SNU-387 and Huh-7) and a human normal hepatocyte line(THLE-2). siRNA was applied to knock down circFBLIM1 and pcDNA vector was used to overexpress circFBLIM1 in SNU-387 cells. Flow cytometry, Cell Counting Kit-8(CCK-8), and wound healing assay were performed to analyze cell apoptosis, proliferation and migration. qPCR and Western blot were adopted to detect the influences of circFBLIM1 knockdown and overexpression on the expressions of miR-338 and PFKL in SNU-387 cells. Results circFBLIM1 was highly expressed in SNU-387 and Huh-7 cells(both P<0.01). Silencing circFBLIM1 in SNU-387 cells significantly inhibited cell proliferation, promoted apoptosis, reduced migration ability, up-regulated miR-338 expression, and down-regulated PFKL mRNA and protein expressions(all P<0.05). Overexpressing circFBLIM1 promoted cell proliferation, enhanced migration ability, down-regulated miR-338, and up-regulated PFKL mRNA and protein expressions(all P<0.05). Conclusions circFBLIM1 is highly expressed in liver cancer cells. circFBLIM1 facilitates the proliferation and migration of liver cancer cells and exerts its effects in liver cancer progression by activating the circFBLIM1/miR-338/PFKL axis in the glycolytic pathway.