Ren Zhijing, Yang Qinqin, Guo Jiajia, Huang Haifeng, Li Bo, Yang Zhen, Tian Xiaobin
Medical College of Guizhou University, Guiyang, China.
Department of Clinical Laboratory, Guizhou Provincial People's Hospital, Guiyang, China.
Front Cell Dev Biol. 2021 Sep 9;9:714601. doi: 10.3389/fcell.2021.714601. eCollection 2021.
An increasing number of studies have demonstrated that circular RNAs (circRNAs) are involved in tumor progression. However, the role of hsa_circ_0000073 in osteosarcoma (OS) is still not fully elucidated. Quantitative reverse transcription-polymerase chain reaction or Western blot was used to detect the gene expression. GeneChip analysis, bioinformatics, luciferase reporter, and RNA immunoprecipitation assays were adopted to predict and verify the relationships between genes. Counting Kit-8 Assay, clone formation assay, wound-healing assay, transwell assays, cell cycle assays, and tumorigenesis were used to evaluate cell function. hsa_circ_0000073 was highly expressed in OS cell lines and could promote OS progression, including proliferation, migration, invasion, and cell cycle as well as tumorigenesis . Mechanically, hsa_circ_0000073 could readily downregulate the expression of CCNE2 and MDM2 through miR-1252-5p. Rescue experiments validated miR-1252-5p mimics, or CCNE2/MDM2 short hairpin RNA could reverse the hsa_circ_0000073 overexpressing-induced impairment of malignant tumor behavior. hsa_circ_0000073 functions as a tumor promoter in OS to increase malignant tumor behavior through sponging miR-1252-5p and regulating CCNE2 and MDM2 expression, which could be a novel target for OS therapy.
越来越多的研究表明，环状RNA（circRNA）参与肿瘤进展。然而，hsa_circ_0000073在骨肉瘤（OS）中的作用仍未完全阐明。采用定量逆转录-聚合酶链反应或蛋白质免疫印迹法检测基因表达。采用基因芯片分析、生物信息学、荧光素酶报告基因和RNA免疫沉淀试验预测和验证基因之间的关系。采用细胞增殖检测试剂盒-8法、克隆形成试验、伤口愈合试验、Transwell试验、细胞周期试验和肿瘤发生试验评估细胞功能。hsa_circ_0000073在OS细胞系中高表达，可促进OS进展，包括增殖、迁移、侵袭、细胞周期以及肿瘤发生。机制上，hsa_circ_0000073可通过miR-1252-5p下调CCNE2和MDM2的表达。挽救实验证实，miR-1252-5p模拟物或CCNE2/MDM2短发夹RNA可逆转hsa_circ_0000073过表达诱导的恶性肿瘤行为损伤。hsa_circ_0000073在OS中作为肿瘤促进因子，通过海绵化miR-1252-5p并调节CCNE2和MDM2的表达来增加恶性肿瘤行为，这可能是OS治疗的一个新靶点。
