Meng Xianbing, Tian Hailong, Guo Wenqiang, Wang Zhigang
Department of Neurosurgery, Qilu Hospital of Shandong University, No. 758, Hefei Road, Shibei District, Qingdao City, Shandong Province, 266035, China.
Department of Neurosurgery, The Second Affiliated Hospital of Shandong First Medical University, Tai'an, 271000, China.
Transl Neurosci. 2021 Nov 19;12(1):456-468. doi: 10.1515/tnsci-2020-0181. eCollection 2021 Jan 1.
Circular RNAs contribute to the progression of glioma. However, the biological role and underlying mechanism of circ_0082375 in glioma remain unclear. Quantitative real-time PCR and Western blot assay were used to evaluate the expression levels of circ_0082375, microRNA-485-5p, and Wnt family member 7B (Wnt7B). The overall survival of glioma patients was estimated by the Kaplan-Meier curve. Cell proliferation, apoptosis, invasion, and migration were detected by cell counting kit-8, 5-ethynyl-2 -deoxyuridine (EdU) staining, flow cytometry, and transwell assays, respectively. Glucose level and lactate production were determined using glucose and lactate assay kits. angiogenesis assay was used to evaluate the angiogenesis of glioma cells. The interaction between microRNA (miR)-485-5p and circ_0082375 or Wnt family member 7B (Wnt7B) was verified by dual-luciferase reporter and RNA immunoprecipitation assays. A xenograft model was used to verify the function of circ_0082375 . circ_0082375 was upregulated in glioma tissues, and it was closely related to the prognosis of glioma patients. circ_0082375 knockdown suppressed cell proliferation, migration, invasion, angiogenesis, glycolysis, and epithelial-mesenchymal transition (EMT), and promoted cell apoptosis in glioma cells. irc_0082375 was a sponge of miR-485-5p, which directly targeted Wnt7B. Knockdown of circ_0082375 inhibited the malignancy, angiogenesis, and glycolysis of glioma cells by sponging miR-485-5p. Besides, circ_0082375 knockdown hampered the growth of glioma growth by regulating the miR-485-5p/Wnt7B axis Altogether, circ_0082375 regulated miR-485-5p/Wnt7B axis to promote the malignancy, angiogenesis, and glycolysis of glioma cells, thereby contributing to the progression of glioma.
环状RNA促进胶质瘤进展。然而,circ_0082375在胶质瘤中的生物学作用及潜在机制仍不清楚。采用定量实时PCR和蛋白质免疫印迹法评估circ_0082375、微小RNA-485-5p和Wnt家族成员7B(Wnt7B)的表达水平。通过Kaplan-Meier曲线评估胶质瘤患者的总生存期。分别采用细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷(EdU)染色、流式细胞术和Transwell实验检测细胞增殖、凋亡、侵袭和迁移。使用葡萄糖和乳酸检测试剂盒测定葡萄糖水平和乳酸生成量。采用血管生成实验评估胶质瘤细胞的血管生成情况。通过双荧光素酶报告基因实验和RNA免疫沉淀实验验证微小RNA(miR)-485-5p与circ_0082375或Wnt家族成员7B(Wnt7B)之间的相互作用。采用异种移植模型验证circ_0082375的功能。circ_0082375在胶质瘤组织中上调,且与胶质瘤患者的预后密切相关。敲低circ_0082375可抑制胶质瘤细胞的增殖、迁移、侵袭、血管生成、糖酵解以及上皮-间质转化(EMT),并促进细胞凋亡。circ_0082375是miR-485-5p的海绵,直接靶向Wnt7B。敲低circ_0082375通过吸附miR-485-5p抑制胶质瘤细胞的恶性程度、血管生成和糖酵解。此外,敲低circ_0082375通过调节miR-485-5p/Wnt7B轴阻碍胶质瘤的生长。总之,circ_0082375通过调节miR-485-5p/Wnt7B轴促进胶质瘤细胞的恶性程度、血管生成和糖酵解,从而推动胶质瘤进展。