Deng Liyong, Gong Kuiyu, Wang Guihua
Department of Oncology, Changsha Central Hospital, No. 161, Shaoshan South Road, Changsha, Hunan, China.
Neurotox Res. 2022 Jun;40(3):825-836. doi: 10.1007/s12640-022-00504-8. Epub 2022 Apr 8.
Hsa_circRNA_0008344 (circ_0008344) is a new glioma-related circular RNA. Our study aims to explore its functions in glioma tumor progression. Real-time quantitative PCR and western blotting were used to detect RNA and protein abundances. RNase R assay, actinomycin D assay, and subcellular fractionation method were performed to identify the features of circ_0008344. Cell-counting kit-8, 5-ethynyl-2'-deoxyuridine assays, transwell assays, tube formation assay, flow cytometry, and nude mice xenograft tumor model were performed. Target relationship was predicted by bioinformatics algorithms and confirmed by dual-luciferase reporter assay. Abundances of circ_0008344 and SUZ RNA binding domain containing 1 (SZRD1) were highly elevated, while miR-638 was downregulated in glioma tumors and cells. Circ_0008344 was identified as a stable circRNA with a circular structure. Silencing circ_0008344 could restrain glioma proliferation, migration, invasion, and angiogenesis. Circ_0008344 functioned as a sponge for miR-638. The negative regulation of circ_0008344 knockdown on glioma progression and angiogenesis could be reversed by miR-638 inhibitor. SZRD1 was a target of miR-318, and its overexpression overturned the inhibition effect of miR-638 mimic on glioma progression and angiogenesis. Meanwhile, we confirmed that circ_0008344 knockdown inhibited SZRD1 expression, and its effect was reversed by miR-638 inhibitor. Also, circ_00008344 knockdown suppressed glioma tumor growth. Circ_0008344 might contribute to glioma progression through miR-638/SZRD1 axis, which might be a novel pathology and treatment target in glioma.
人源环状RNA_0008344(circ_0008344)是一种新的与胶质瘤相关的环状RNA。我们的研究旨在探讨其在胶质瘤肿瘤进展中的作用。采用实时定量PCR和蛋白质免疫印迹法检测RNA和蛋白质丰度。进行了核糖核酸酶R检测、放线菌素D检测和亚细胞分级分离法以鉴定circ_0008344的特征。进行了细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷检测、Transwell检测、管形成检测、流式细胞术和裸鼠异种移植瘤模型实验。通过生物信息学算法预测靶标关系,并通过双荧光素酶报告基因检测进行验证。circ_0008344和含SUZ RNA结合结构域1(SZRD1)的丰度在胶质瘤肿瘤和细胞中显著升高,而miR-638表达下调。circ_0008344被鉴定为具有环状结构的稳定环状RNA。沉默circ_0008344可抑制胶质瘤的增殖、迁移、侵袭和血管生成。circ_0008344作为miR-638的海绵发挥作用。miR-638抑制剂可逆转circ_000