Wang Xinfang, Xu Dong, Pei Xin, Zhang Yingying, Zhang Yuling, Gu Yaxing, Li Ying
Department of Pediatrics, Shandong Provincial Western Hospital, Jinan 250022, Shandong, People's Republic of China.
Cancer Manag Res. 2020 Dec 30;12:13415-13426. doi: 10.2147/CMAR.S272753. eCollection 2020.
Medulloblastoma (MB) is the most common malignant brain tumor during childhood. Circular RNA (circSKA3) was identified to function as an oncogene in MB. However, the mechanism of circSKA3 in MB remains unclear.
The levels of circSKA3, microRNA-383-5p (miR-383-5p), and forkhead box M1 () in MB tissues were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The cell viability and apoptotic rate were assessed via 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and flow cytometry, respectively. The protein levels of B-cell lymphoma 2 (Bcl-2), C-Caspase3, and FOXM1 were detected via Western blot assay. Cell cycle was detected by flow cytometry. The migration and invasion abilities were monitored by Transwell assay. The dual-luciferase reporter assay was constructed to verify the interactions between miR-383-5p and circSKA3 or . The mice model experiment was carried out to validate the effects of circSKA3 in vivo.
The levels of circSKA3 and were significantly elevated, while the level of miR-383-5p was notably declined in MB tissues. CircSKA3 was validated to sponge miR-383-5p, and was a candidate target of miR-383-5p. CircSKA3 silencing impeded cell proliferation, migration, and invasion while promoted apoptosis by targeting miR-383-5p in vitro and retarded xenograft tumor growth in vivo. miR-383-5p suppressed cell proliferation, migration, and invasion but promoted apoptosis in MB cells by regulating FOXM1. CircSKA3 depletion decreased expression via miR-383-5p in MB cells.
CircSKA3 augmented MB progression partly through miR-383-5p/ axis.
髓母细胞瘤（MB）是儿童期最常见的恶性脑肿瘤。环状RNA（circSKA3）被鉴定为在MB中起癌基因作用。然而，circSKA3在MB中的作用机制仍不清楚。
采用定量实时聚合酶链反应（qRT-PCR）检测MB组织中circSKA3、微小RNA-383-5p（miR-383-5p）和叉头框M1（FOXM1）的水平。分别通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐（MTT）法和流式细胞术评估细胞活力和凋亡率。通过蛋白质免疫印迹法检测B细胞淋巴瘤2（Bcl-2）、半胱天冬酶3（C-Caspase3）和FOXM1的蛋白质水平。通过流式细胞术检测细胞周期。通过Transwell法监测迁移和侵袭能力。构建双荧光素酶报告基因检测以验证miR-383-5p与circSKA3或FOXM1之间的相互作用。进行小鼠模型实验以验证circSKA3在体内的作用。
在MB组织中，circSKA3和FOXM1水平显著升高，而miR-383-5p水平显著下降。验证circSKA3可吸附miR-383-5p，且FOXM1是miR-383-5p的候选靶标。在体外，circSKA3沉默通过靶向miR-383-5p抑制细胞增殖、迁移和侵袭，同时促进细胞凋亡，在体内则抑制异种移植肿瘤生长。miR-383-5p通过调节FOXM1抑制MB细胞的增殖、迁移和侵袭，但促进细胞凋亡。在MB细胞中，circSKA3缺失通过miR-383-5p降低FOXM1表达。
CircSKA3部分通过miR-383-5p/FOXM1轴促进MB进展。
