Circular RNA hsa_circ_0000073 Enhances Osteosarcoma Cells Malignant Behavior by Sponging miR-1252-5p and Modulating CCNE2 and MDM2.
Circular RNA hsa_circ_0000073 Enhances Osteosarcoma Cells Malignant Behavior by Sponging miR-1252-5p and Modulating CCNE2 and MDM2.
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DOI：
10.3389/fcell.2021.714601
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发表时间：
2021
影响因子：
5.5
通讯作者：
Tian X
中科院分区：
文献类型：
--
作者：
Ren Z;Yang Q;Guo J;Huang H;Li B;Yang Z;Tian X
目的：越来越多的研究表明，环状RNA（circular RNA，circRNA）参与了肿瘤的发生、发展过程。然而，hsa_circ_0000073在骨肉瘤（OS）中的作用仍未完全阐明。方法：采用定量逆转录-聚合酶链反应（RT-PCR）或Western blot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免疫沉淀技术预测和验证基因间的关系。使用计数试剂盒-8测定、克隆形成测定、伤口愈合测定、transwell测定、细胞周期测定和体内肿瘤发生来评价细胞功能。结果如下：hsa_circ_0000073在骨肉瘤细胞株中高表达，在体外可促进骨肉瘤的增殖、迁移、侵袭、细胞周期和体内成瘤。hsa_circ_0000073可通过miR-1252- 5 p下调CCNE 2和MDM 2的表达。挽救实验验证了miR-1252- 5 p模拟物或CCNE 2/MDM 2短发夹RNA可以逆转hsa_circ_0000073过表达诱导的恶性肿瘤行为损害。总结：hsa_circ_0000073在OS中作为肿瘤启动子，通过海绵状作用于miR-1252- 5 p，调控CCNE 2和MDM 2的表达，增加恶性肿瘤的发生，有望成为OS治疗的新靶点。
Objective: 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. Methods: 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 in vivo tumorigenesis were used to evaluate cell function. Results: hsa_circ_0000073 was highly expressed in OS cell lines and could promote OS progression, including proliferation, migration, invasion, and cell cycle in vitro as well as tumorigenesis in vivo. 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. Conclusion: 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.
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