Huo Shousong, Dou Dongmei
Department of Orthopaedic, Huaihe Hospital of Henan University, Kaifeng, 475000, Henan, People's Republic of China.
Institute of Chronic Disease Risk Assessment, Henan University, Jinming Campus, Kaifeng, 475000, Henan, People's Republic of China.
Cancer Manag Res. 2021 Feb 10;13:1257-1270. doi: 10.2147/CMAR.S290645. eCollection 2021.
Osteosarcoma (OS) is a common malignant bone cancer that occurs in adolescents and children. Circular RNAs (circRNAs) are important regulators of tumorigenesis and development. This study aimed to explore the role and molecular basis of circ_0056285 in OS.
The levels of circ_0056285, miR-1244 and tripartite motif containing 44 (TRIM44) were determined by quantitative real-time polymerase chain reaction or Western blot assay. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8) assay and colony formation assay. Cell apoptosis was assessed by flow cytometry and caspase 3and caspase 9 activity assay kits. Glucose uptake, lactate product and ATP level were examined using commercial kits. Hexokinase II (HK2) and lactate dehydrogenase A (LDHA) levels were measured by Western blot assay. The interaction among circ_0056285, miR-1244 and TRIM44 was confirmed by dual-luciferase reporter assay, RNA immunoprecipitation assay or RNA pull-down assay. Xenograft experiment was conducted to explore tumor growth in vivo. Exosomes were identified by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA) and Western blot. The diagnostic value of exosomal circ_0056285 was evaluated by receiver operating characteristic (ROC) curve.
Circ_0056285 and TRIM44 were up-regulated, and miR-1244 was down-regulated in OS tissues and cells. Circ_0056285 silencing inhibited proliferation and glycolysis and promoted apoptosis in OS cells. Also, circ_0056285 knockdown hindered proliferation and accelerated apoptosis in OS cells by regulating miR-1244/TRIM44 axis. Circ_0056285 depletion impeded tumor growth in vivo. Furthermore, ROC curve showed that circ_0056285 might be a diagnostic biomarker in OS.
Circ_0056285 facilitated OS progression by sponging miR-1244 and increasing TRIM44 expression, providing a promising therapeutic target for OS.
骨肉瘤（OS）是一种常见于青少年和儿童的恶性骨癌。环状RNA（circRNAs）是肿瘤发生和发展的重要调节因子。本研究旨在探讨circ_0056285在骨肉瘤中的作用及分子机制。
采用定量实时聚合酶链反应或蛋白质免疫印迹法检测circ_0056285、miR-1244和含三联基序蛋白44（TRIM44）的表达水平。通过细胞计数试剂盒-8（CCK-8）法和集落形成试验评估细胞增殖。采用流式细胞术和半胱天冬酶3及半胱天冬酶9活性检测试剂盒评估细胞凋亡。使用商业试剂盒检测葡萄糖摄取、乳酸生成和ATP水平。通过蛋白质免疫印迹法检测己糖激酶II（HK2）和乳酸脱氢酶A（LDHA）水平。采用双荧光素酶报告基因检测、RNA免疫沉淀试验或RNA下拉试验证实circ_0056285、miR-1244和TRIM44之间的相互作用。进行异种移植实验以探讨体内肿瘤生长情况。通过透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和蛋白质免疫印迹法鉴定外泌体。采用受试者工作特征（ROC）曲线评估外泌体circ_0056285的诊断价值。
在骨肉瘤组织和细胞中，circ_0056285和TRIM44表达上调，miR-1244表达下调。circ_0056285沉默抑制骨肉瘤细胞增殖和糖酵解并促进细胞凋亡。此外，circ_0056285敲低通过调节miR-1244/TRIM44轴阻碍骨肉瘤细胞增殖并加速细胞凋亡。circ_0056285缺失抑制体内肿瘤生长。此外，ROC曲线显示circ_0056285可能是骨肉瘤的诊断生物标志物。
circ_0056285通过吸附miR-1244并增加TRIM44表达促进骨肉瘤进展，为骨肉瘤提供了一个有前景的治疗靶点。
