人源环状RNA hsa_circ_0000006通过靶向骨肉瘤中的免疫球蛋白样结构域蛋白1（LRIG1）的miR-361-3p诱导肿瘤发生。
hsa_circ_0000006 induces tumorigenesis through miR-361-3p targeting immunoglobulin-like domains protein 1 (LRIG1) in osteosarcoma.
作者信息
Gao Yang, Liu Chengtao, Zhao Xiaoling, Liu Chaojun, Bi Wenzhi, Jia Jinpeng
机构信息
Department of Orthopaedics, General Hospital of Chinese People's Liberation Army, Beijing, China.
Shandong Wendeng Osteopathic Hospital, Weihai, China.
出版信息
Ann Transl Med. 2021 Aug;9(15):1242. doi: 10.21037/atm-21-3076.
BACKGROUND
Osteosarcoma (OS) is considered to be the most highly prevalent bone tumor. In the progression of different human cancers, the role of circular RNAs (circRNAs) has been extensively studied. Microarray analysis has indicated that hsa_circ_0000006 expression was lower in OS, but the mechanism of hsa_circ_0000006 in regulating the progression of OS remains elusive.
METHODS
The expression of cancer-related genes at the transcriptional and translational levels was assessed by RT-qPCR and western blotting (WB). Colony formation and Cell Counting Kit-8 (CCK-8) assays were used to evaluate the proliferative potential of cells. The transwell assay was used to examine the invasive and migratory potential of cells. Furthermore, dual-luciferase reporter (DLR) and RNA pull-down assays were performed for the validation of the targeting sites of hsa_circ_0000006, miR-361-3p, and the 3'-untranslated region (3'-UTR) of immunoglobulin-like domains protein 1 (LRIG1) mRNA. Moreover, the protein levels of epithelial-to-mesenchymal transition (EMT) markers were analyzed by WB.
RESULTS
The expression of hsa_circ_0000006 and LRIG1 were found to be down-regulated in OS tissues and cells, while miR-361-3p was up-regulated. Knockdown of hsa_circ_0000006 promoted the progression and development of OS, as well as EMT. Furthermore, hsa_circ_0000006 was revealed as a sponge of miR-361-3p, which negatively regulates miR-361-3p expression. LRIG1 was found to be an miR-361-3p target. In OS cells, the LRIG1 expression level was decreased, with elevated expression of miR-361-3p. Advanced studies demonstrated that hsa_circ_0000006 regulates LRIG1 expression through sponging miR-361-3p, then promotes the tumorigenesis of OS.
CONCLUSIONS
hsa_circ_0000006 is associated with the progression and development of OS through miR-361-3p by target LRIG1, which is a significant biomarker and effective therapeutic target for patients with OS.
背景
骨肉瘤（OS）被认为是最常见的骨肿瘤。在不同人类癌症的进展过程中，环状RNA（circRNAs）的作用已得到广泛研究。微阵列分析表明，hsa_circ_0000006在骨肉瘤中的表达较低，但hsa_circ_0000006调控骨肉瘤进展的机制仍不清楚。
方法
通过逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法（WB）评估癌症相关基因在转录和翻译水平的表达。采用集落形成实验和细胞计数试剂盒-8（CCK-8）实验评估细胞的增殖潜力。采用Transwell实验检测细胞的侵袭和迁移潜力。此外，进行双荧光素酶报告基因（DLR）和RNA下拉实验，以验证hsa_circ_0000006、miR-361-3p和免疫球蛋白样结构域蛋白1（LRIG1）mRNA的3'-非翻译区（3'-UTR）的靶向位点。此外，通过WB分析上皮-间质转化（EMT）标志物的蛋白水平。
结果
发现hsa_circ_0000006和LRIG1在骨肉瘤组织和细胞中的表达下调，而miR-361-3p上调。敲低hsa_circ_0000006促进了骨肉瘤的进展和发展以及EMT。此外，hsa_circ_0000006被发现是miR-361-3p的海绵，负向调节miR-361-3p的表达。LRIG1被发现是miR-361-3p的靶标。在骨肉瘤细胞中，LRIG蛋白表达水平降低，miR-361-3p表达升高。进一步研究表明，hsa_circ_0000006通过吸附miR-361-3p调节LRIG1表达，进而促进骨肉瘤的肿瘤发生。
结论
hsa_circ_0000006通过靶向LRIG1与miR-361-3p相关，参与骨肉瘤的进展和发展，LRIG1是骨肉瘤患者的重要生物标志物和有效的治疗靶点。
Zotero 插件