Lu Jun, Lou Gaojie, Jiang Lin, Liu Xiaoxing, Jiang Jianxin, Wang Xiaolin
Department of Neurosurgery, Taizhou People's Hospital, Taizhou, China.
Front Neurol. 2021 Nov 18;12:679745. doi: 10.3389/fneur.2021.679745. eCollection 2021.
Circular RNA (circNUP98) has been reported to promote renal cancer; however, its role in other cancers is unknown. The function of circNUP98 in glioblastoma (GB) cancer was explored in this study. A total of 58 GB tissue samples were collected to study the expression of circNUP98 and miR-519a-3p [both the mature and pre-mature microRNA (miRNA)] by quantitative real-time PCR (RT-qPCR) and heatmap analysis. The subcellular location that expresses circNUP98 was analyzed by nuclear fractionation assay. RNA pull-down assay was performed to evaluate the interaction between circNUP98 and pre-mature miR-519a-3p. Overexpression assays were performed to investigate the role of circNUP98 in the regulation of both the mature and pre-mature miR-519a-3p. The role of circNUP98 and miR-519a-3p in GB cell proliferation was explored by 5-bromo-2-deoxyuridine (BrdU) assay and was assessed in mouse xenograft model. Heatmap analysis showed that circNUP98 and pre-mature miR-519a-3p were upregulated in GB, while mature miR-519a-3p was downregulated in GB. Across the cancer tissues, circNUP98 was inversely correlated with mature miR-519a-3p, but positively correlated with pre-mature miR-519a-3p. In GB cells, circNUP98 was localized to both the nucleus and cytoplasm and it interacted with pre-mature miR-519a-3p. In GB cells, circNUP98 increased the expression levels of pre-mature miR-519a-3p and decreased the expression levels of mature miR-519a-3p. BrdU and cholecystokinin octapeptide (CCK-8) assays illustrated that overexpression of circNUP98 reduced the inhibitory effects of miR-519a-3p on cell proliferation. CircNUP98 contributed to larger tumors, which resulted in significantly reduced mice survival. CircNUP98 suppresses the maturation of miR-519a-3p to promote GB cell proliferation.
据报道,环状RNA(circNUP98)可促进肾癌;然而,其在其他癌症中的作用尚不清楚。本研究探讨了circNUP98在胶质母细胞瘤(GB)中的功能。共收集58份GB组织样本,通过定量实时聚合酶链反应(RT-qPCR)和热图分析研究circNUP98和miR-519a-3p[成熟和前体微小RNA(miRNA)]的表达。通过细胞核分级分离试验分析表达circNUP98的亚细胞定位。进行RNA下拉试验以评估circNUP98与前体miR-519a-3p之间的相互作用。进行过表达试验以研究circNUP98在调节成熟和前体miR-519a-3p中的作用。通过5-溴-2-脱氧尿苷(BrdU)试验探索circNUP98和miR-519a-3p在GB细胞增殖中的作用,并在小鼠异种移植模型中进行评估。热图分析显示,circNUP98和前体miR-519a-3p在GB中上调,而成熟miR-519a-3p在GB中下调。在所有癌症组织中,circNUP98与成熟miR-519a-3p呈负相关,但与前体miR-519a-3p呈正相关。在GB细胞中,circNUP98定位于细胞核和细胞质,并与前体miR-519a-3p相互作用。在GB细胞中,circNUP98增加前体miR-519a-3p的表达水平,并降低成熟miR-519a-3p的表达水平。BrdU和胆囊收缩素八肽(CCK-8)试验表明,circNUP98的过表达降低了miR-519a-3p对细胞增殖的抑制作用。CircNUP98导致肿瘤更大,从而显著降低小鼠存活率。CircNUP98抑制miR-519a-3p的成熟以促进GB细胞增殖。