Wang Bin, Li Bing, Si Tongguo
Department of Interventional Oncology, Tianjin Huanhu Hospital (Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases, Department of Interventional Therapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer), Tianjin 300350, China.
Department of Neurosurgery, Tianjin Huanhu Hospital (Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases), Tianjin 300350, China.
Brain Res. 2020 Dec 1;1748:147108. doi: 10.1016/j.brainres.2020.147108. Epub 2020 Sep 4.
Circular RNAs (circRNAs) play important roles in the development and treatment of glioma. However, the role and mechanism of circRNA carboxypeptidase A4 (circ0082374) in glioma are largely unknown. Forty-two glioma patients and 28 normal patients were recruited. Glioma cell lines A172 and U251 were used for functional assays. The expression levels of circ0082374, microRNA-326 (miR-326) and sirtuin 1 (SIRT1) were examined via quantitative real-time polymerase chain reaction or western blot. Cell viability, migration, invasion and glycolysis were measured via cell counting kit-8, trans-well, oxygen consumption rate and western blot, respectively. The target correlation of circ0082374/miR-326 or miR-326/SIRT1 was explored via dual-luciferase reporter, RNA immunoprecipitation and pull-down assays. The role of circ0082374 in vivo was investigated via xenograft model. We found circ0082374 expression was elevated in glioma tissues and cells. Knockdown of circ0082374 suppressed the viability, migration, invasion and glycolysis in glioma cells. miR-326 was a target of circ0082374 and miR-326 knockdown attenuated the inhibitive role of circ0082374 silence in glioma progression. SIRT1 was a target of miR-326 and circ0082374 could promote SIRT1 expression by sponging miR-326. Silence of SIRT1 reversed the promoting effect of circ0082374 on glioma progression. Knockdown of circ0082374 reduced xenograft tumor growth by miR-326/SIRT1 in vivo. Collectively, silence of circ0082374 repressed the viability, migration, invasion and glycolysis in glioma cells by regulating miR-326 and SIRT1 in a ceRNA mechanism, providing a new mechanism for the pathogenesis of glioma.
环状RNA（circRNAs）在胶质瘤的发生发展及治疗中发挥着重要作用。然而，circRNA羧肽酶A4（circ0082374）在胶质瘤中的作用及机制尚不清楚。本研究招募了42例胶质瘤患者和28例正常对照者。采用胶质瘤细胞系A172和U251进行功能实验。通过定量实时聚合酶链反应或蛋白质免疫印迹法检测circ0082374﹑微小RNA-326（miR-326）和沉默调节蛋白1（SIRT1）的表达水平。分别采用细胞计数试剂盒-8法、Transwell小室法、耗氧率检测法及蛋白质免疫印迹法检测细胞活力、迁移、侵袭及糖酵解水平。通过双荧光素酶报告基因实验、RNA免疫沉淀实验及下拉实验探究circ0082374/miR-326或miR-326/SIRT1的靶向关系。通过异种移植模型研究circ0082374在体内的作用。我们发现circ0082374在胶质瘤组织和细胞中表达上调。敲低circ0082374可抑制胶质瘤细胞的活力、迁移、侵袭及糖酵解。miR-326是circ0082374的靶标，敲低miR-326可减弱circ0082374沉默对胶质瘤进展的抑制作用。SIRT1是miR-326的靶标，circ0082374可通过吸附miR-326促进SIRT1表达。沉默SIRT1可逆转circ0082374对胶质瘤进展的促进作用。在体内，敲低circ0082374可通过miR-326/SIRT1途径抑制异种移植瘤的生长。总之，沉默circ0082374可通过ceRNA机制调控miR-326和SIRT1，从而抑制胶质瘤细胞的活力、迁移、侵袭及糖酵解，为胶质瘤的发病机制提供了新的理论依据。
