背景:胶质瘤是主要的恶性脑肿瘤。这项研究试图阐明糖酵解相关的lncARSR对神经胶质瘤的影响和前瞻性机制。
方法:在正常神经胶质细胞和神经胶质瘤细胞中评估LncARSR水平。细胞增殖,迁移,通过CCK-8,伤口愈合,和transwell分析。流式细胞术用于测量细胞凋亡和细胞周期。采用生化试剂盒和免疫印迹法检测糖酵解相关指标的含量和蛋白表达,分别。我们使用双荧光素酶报告测定和染色质免疫沉淀(ChIP)实验分析了lncARSR敲低和信号转导和转录激活因子3(STAT3)的过表达对己糖激酶2(HK2)的影响。通过动物实验进一步评估lncARSR对神经胶质瘤进展的影响。
结果:与正常神经胶质细胞相比,LncARSR在神经胶质瘤细胞中的表达水平升高。过表达lncARSR增强了增殖,迁移,入侵,和G2/M期阻滞在神经胶质瘤细胞,也增加了葡萄糖,乳酸,ATP生产,以及HK2、PFK1、PKM2、GLUT1和LDHA的表达。抑制lncARSR后,STAT3与HK2基因启动子的结合减弱。上调STAT3逆转了敲低lncARSR对细胞增殖的抑制功能,迁移,入侵,G2/M阶段停止,和糖酵解,并抵消其对细胞凋亡的促进作用。在体内,敲低lncARSR抑制胶质瘤生长和ki67和PCNA表达。
结论:LncARSR通过STAT3-HK2轴促进糖酵解促进胶质瘤的发展。
BACKGROUND: Glioma represents the predominant malignant brain tumor. This investigation endeavors to elucidate the impact and prospective mechanisms of glycolysis-related lncARSR on glioma.
METHODS: LncARSR level was assessed in normal glial cells and glioma cells. Cell proliferation, migration, and invasion measurements were conducted through CCK-8, wound healing, and transwell assay. Flow cytometry was utilized to measure cell apoptosis and cell cycle. Biochemical assay kits and immunoblotting were employed to measure the content of glycolysis-related indicators and protein expression, respectively. We analyzed the impact of both lncARSR knockdown and overexpression of the Signal Transducer and Activator of Transcription 3 (STAT3) on
Hexokinase 2 (HK2) using dual luciferase reporter assays and Chromatin Immunoprecipitation (ChIP) experiments. Further assessment of the impact of lncARSR on glioma progression was conducted through animal experiments.
RESULTS: LncARSR was expressed at elevated levels in glioma cells compared to normal glial cells. Overexpressing lncARSR enhanced proliferation, migration, invasion, and G2/M phase arrest in glioma cells and also increased glucose, lactate, ATP production, as well as the expression of HK2, PFK1, PKM2, GLUT1, and LDHA. STAT3 binding to the HK2 gene promoter was weakened following the knockdown of lncARSR. Upregulation of STAT3 reversed the suppressed functions of knocking down lncARSR on cell proliferation, migration, invasion, G2/M phase arrest, and glycolysis and counteracted its promotional effect on cell apoptosis. In vivo, knocking down lncARSR inhibits glioma growth and ki67 and PCNA expression.
CONCLUSIONS: LncARSR promotes the development of glioma by enhancing glycolysis through the STAT3-HK2 axis.