Li Fubin, Wu Lin, Liu Bin, An Xiangyang, Du Xinrui
Department of Neurosurgery, Zibo Central Hospital Zibo 255036, Shandong, China.
Department of Pediatrics, Zhangdian Maternal and Child Health Care Hospital Zibo 255036, Shandong, China.
Am J Cancer Res. 2023 Jul 15;13(7):2906-2921. eCollection 2023.
Recent studies have verified that circRNAs (circular RNAs) play a critical role in glioma occurrence and malignant progression. However, numerous circRNAs with unknown functions remain to be explored with further research. qPCR (quantitative real-time polymerase chain reaction) was employed to detect circTIE1 expression in glioma tissues, NHAs (normal human astrocytes), and glioma cellular lines (U87, U118, U251, T98G, LN229). Cell viability was evaluated by CCK-8 assay. Cellular proliferation was evaluated by a 5-ethynyl-2'-deoxyuridine (EdU) proliferation assay. Cell migration and aggression were both evaluated by transwell and migration assays. The direct binding and regulation among circTIE1, miR-1286 and TEAD1 was identified by western blotting, qPCR, luciferase reporter assay, and RNA immunoprecipitation (RIP) assay. Xenografts were generated by injecting glioma cells orthotopically into the brains of nude mice. Immunohistochemistry staining was implemented to evaluate the expression of the proliferation markers ki67 and TEAD1. We found that circTIE1 (circBase ID: hsa_circ_0012012) was upregulated in glioma tissues and glioma cellular lines in contrast to NBT (normal brain tissues) and NHA. CircTIE1 knockdown inhibited glioma cell viability, proliferation, migration and aggression both in vitro and in vivo. Mechanistically, circTIE1 could upregulate TEAD1 expression via miR-1286 sponging, and TEAD1 is a well-known functional gene that could promote malignant advancement in glioma. This research found a novel circRNA, circTIE1, which is an essential marker of glioma progression and diagnosis and may be anticipated to become a crucial target for molecular targeted therapy of glioma.
最近的研究证实，环状RNA（circRNAs）在胶质瘤的发生和恶性进展中起关键作用。然而，众多功能未知的环状RNA仍有待进一步研究探索。采用qPCR（定量实时聚合酶链反应）检测胶质瘤组织、正常人星形胶质细胞（NHAs）和胶质瘤细胞系（U87、U118、U251、T98G、LN229）中circTIE1的表达。通过CCK-8法评估细胞活力。通过5-乙炔基-2'-脱氧尿苷（EdU）增殖试验评估细胞增殖。通过Transwell和迁移试验评估细胞迁移和侵袭能力。通过蛋白质免疫印迹法、qPCR、荧光素酶报告基因试验和RNA免疫沉淀（RIP）试验确定circTIE1、miR-1286和TEAD1之间的直接结合和调控关系。将胶质瘤细胞原位注射到裸鼠脑内构建异种移植瘤模型。采用免疫组织化学染色评估增殖标志物ki67和TEAD1的表达。我们发现，与正常脑组织（NBT）和正常人星形胶质细胞相比，circTIE1（circBase ID：hsa_circ_0012012）在胶质瘤组织和胶质瘤细胞系中表达上调。敲低circTIE1可在体外和体内抑制胶质瘤细胞的活力、增殖、迁移和侵袭能力。机制上，circTIE1可通过海绵吸附miR-1286上调TEAD1的表达，而TEAD1是一个众所周知的功能性基因，可促进胶质瘤的恶性进展。本研究发现了一种新的环状RNA，circTIE1，它是胶质瘤进展和诊断的重要标志物，有望成为胶质瘤分子靶向治疗的关键靶点。
