Song Dewen, Ye Lin, Xu Zengliang, Jin Yanfei, Zhang Lintao
Department of Neurosurgery, Jiaozhou people's Hospital, Qingdao, Shandong, China.
Neoplasma. 2021 Mar;68(2):391-403. doi: 10.4149/neo_2020_200702N682. Epub 2021 Jan 14.
Circular RNAs (circRNAs) play a crucial role in tumor occurrence and progression. And the dysregulated circRNAs are reported to be relevant to glioma development. Nevertheless, the function and regulatory mechanism of hsa_circ_0030018 in glioma progression are largely indistinct. The abundances of hsa_circ_0030018, miR-1297, and RAB21 were detected using quantitative real-time polymerase chain reaction or western blot. Cell proliferation was assessed via colony formation assay and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell apoptosis and cell cycle progression were evaluated by flow cytometry. Cell migration and invasion were examined using transwell assay and wound healing assay. The protein levels were measured by western blot. The interaction between miR-1297 and hsa_circ_0030018 or RAB21 was validated via dual-luciferase reporter analysis, RNA immunoprecipitation (RIP), and RNA pull-down assays. A xenograft model experiment was performed to analyze the function of hsa_circ_0030018 on tumor growth in vivo. hsa_circ_0030018 and RAB21 levels were enhanced, and the miR-1297 level was reduced in glioma tissues and cells. The silence of hsa_circ_0030018 or overexpression of miR-1297 impeded cell proliferation, metastasis, and expedited cell apoptosis and cycle arrest in glioma cells. Furthermore, hsa_circ_0030018 modulated glioma malignant behaviors via sponging miR-1297, and miR-1297 suppressed glioma development via targeting RAB21. Moreover, hsa_circ_0030018 knockdown inhibited tumor growth in vivo. The hsa_circ_0030018 knockdown repressed glioma progression by mediating the miR-1297/RAB21 pathway, providing potential therapeutic targets for glioma treatment.
环状RNA（circRNAs）在肿瘤的发生和发展中起着至关重要的作用。据报道，失调的circRNAs与胶质瘤的发展相关。然而，hsa_circ_0030018在胶质瘤进展中的功能和调控机制在很大程度上尚不清楚。使用定量实时聚合酶链反应或蛋白质免疫印迹法检测hsa_circ_0030018、miR-1297和RAB21的丰度。通过集落形成试验和3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐（MTT）试验评估细胞增殖。通过流式细胞术评估细胞凋亡和细胞周期进程。使用Transwell试验和伤口愈合试验检测细胞迁移和侵袭。通过蛋白质免疫印迹法测量蛋白质水平。通过双荧光素酶报告基因分析、RNA免疫沉淀（RIP）和RNA下拉试验验证miR-1297与hsa_circ_0030018或RAB21之间的相互作用。进行异种移植模型实验以分析hsa_circ_0030018在体内对肿瘤生长的作用。在胶质瘤组织和细胞中，hsa_circ_0030018和RAB21水平升高，而miR-1297水平降低。沉默hsa_circ_0030018或过表达miR-1297可抑制胶质瘤细胞的增殖、转移，并加速细胞凋亡和细胞周期停滞。此外，hsa_circ_0030018通过海绵吸附miR-1297调节胶质瘤的恶性行为，而miR-1297通过靶向RAB21抑制胶质瘤的发展。此外，敲低hsa_circ_0030018可抑制体内肿瘤生长。敲低hsa_circ_0030018通过介导miR-1297/RAB21途径抑制胶质瘤进展，为胶质瘤治疗提供了潜在的治疗靶点。
