Deng Yu, Xu Liu, Li Yuqiang
Department of Pathology, Jinan People's Hospital, No. 001 Xuehu Street, Zhangjiawa Street, Laiwu District, Jinan, Shandong, 271100, China.
Transl Neurosci. 2023 Aug 2;14(1):20220294. doi: 10.1515/tnsci-2022-0294. eCollection 2023 Jan 1.
The effect of circular RNA in many human cancers is widely studied. Nevertheless, their specific biological functions and mechanisms in glioma remain unclear.
CircEXOC6, miR-433-3p, and frizzled class receptor 6 (FZD6) mRNA expression levels were measured by quantitative reverse transcription polymerase chain reaction assay. Cell proliferation, migration, invasion, apoptosis, and angiogenesis were tested by colony formation, cell-light 5-ethynyl-2'-deoxyuridine, transwell, and tube formation assays, respectively. Moreover, glucose consumption and lactate production were calculated to evaluate the glycolytic metabolism using the respective kits. Western blot assay was carried out to measure the protein levels of apoptotic markers (Bcl-2 and Bax), glycolytic markers (HK2 and GLUT1), and FZD6. The targeted relationship of miR-433-3p and circEXOC6 or FZD6 was verified by dual-luciferase reporter or RNA immunoprecipitation assays. , xenograft and immunohistochemistry assay was conducted to discriminate the effect of circEXOC6.
CircEXOC6 and FZD6 were highly expressed, while miR-433-3p was significantly lowly expressed in glioma tissues or cells. Deficiency of circEXOC6 inhibited cell proliferation, migration, invasion, angiogenesis, and glycolysis, and triggered cell apoptosis ratio in glioma; simultaneously, it could block the growth of tumor . In addition, miR-433-3p was a target of circEXOC6, and downregulated miR-433-3p could partly weaken the inhibitory effect of circEXOC6 deficiency. Besides, miR-433-3p enrichment inhibited cell progression and glycolysis in glioma, and the effect was reversed by overexpression of FZD6.
Deletion of circEXOC6 restrained cell progression and glycolysis by sponging miR-433-3p and interacting with FZD6, which might provide an underlying target for glioma treatment.
环状RNA在多种人类癌症中的作用已得到广泛研究。然而,它们在胶质瘤中的具体生物学功能和机制仍不清楚。
采用定量逆转录聚合酶链反应检测法测定环状外切酶6(CircEXOC6)、微小RNA-433-3p(miR-433-3p)和卷曲蛋白家族受体6(FZD6)mRNA的表达水平。分别通过集落形成、细胞轻5-乙炔基-2'-脱氧尿苷、Transwell和管形成试验检测细胞增殖、迁移、侵袭、凋亡和血管生成。此外,使用相应试剂盒计算葡萄糖消耗和乳酸生成以评估糖酵解代谢。进行蛋白质印迹分析以检测凋亡标志物(Bcl-2和Bax)、糖酵解标志物(HK2和GLUT1)和FZD6的蛋白质水平。通过双荧光素酶报告基因或RNA免疫沉淀试验验证miR-433-3p与CircEXOC6或FZD6的靶向关系。进行异种移植和免疫组织化学试验以鉴别CircEXOC6的作用。
CircEXOC6和FZD6在胶质瘤组织或细胞中高表达,而miR-433-3p显著低表达。CircEXOC6缺失抑制胶质瘤细胞增殖、迁移、侵袭、血管生成和糖酵解,并引发细胞凋亡率升高;同时,它可以抑制肿瘤生长。此外,miR-433-3p是CircEXOC6的靶点,下调miR-433-3p可部分减弱CircEXOC6缺失的抑制作用。此外,miR-433-3p富集抑制胶质瘤细胞进展和糖酵解,而FZD6过表达可逆转这种作用。
CircEXOC6缺失通过海绵化miR-433-3p并与FZD6相互作用来抑制细胞进展和糖酵解,这可能为胶质瘤治疗提供潜在靶点。