长链非编码RNA TUG1通过增强ZBTB7C的表达促进骨肉瘤的恶性进展。
Long noncoding RNA TUG1 promotes malignant progression of osteosarcoma by enhancing ZBTB7C expression.
发表日期：2023 Aug 08
作者：
Xueying An, Wenshu Wu, Pu Wang, Abdurahman Mahmut, Junxia Guo, Jian Dong, Wang Gong, Bin Liu, Lin Yang, Yuze Ma, Xingquan Xu, Jianmei Chen, Wangsen Cao, Qing Jiang
来源：
Biomedical Journal
摘要：
长链非编码RNA（lncRNA）的紊乱是肿瘤发生的重要因素。lncRNA牛磺酸上调基因1（lncTUG1）的异常表达在多种肿瘤中已被报道；然而，其在骨肉瘤（OS）进展中的精确作用和关键靶点尚不清楚。通过生物信息学分析评估和OS细胞的qRT-PCR确认，评估了与OS进展相关的lncRNA和其调控的miRNA的表达谱。通过转录组测序鉴定出了miRNA的靶点，并通过荧光素酶报告基因和RNA pull-down实验进行验证。建立了几种体内和体外方法，包括CCK8测定、免疫印迹、qRT-PCR、慢病毒转导和OS细胞异种移植小鼠模型，以验证lncTUG1调控miRNA和下游靶基因对OS细胞的生长、凋亡和进展的影响。我们发现，OS细胞中lncTUG1和miR-26a-5p表达呈相反的上调或下调，lncTUG1的siRNA介导的沉默逆转了miR-26a-5p的下调，并抑制了OS细胞的增殖，增强了其凋亡。此外，我们还发现一个Oncoprotein ZBTB7C在接受lncTUG1/miR-26a-5p调控的OS细胞中也上调。更重要的是，ZBTB7C的沉默减少了ZBTB7C的上调，而ZBTB7C的过表达减弱了lncTUG1沉默对OS移植瘤模型的抗癌作用。我们的数据表明，lncTUG1作为miR-26a-5p的海绵，通过上调ZBTB7C促进了OS的进展，靶向lncTUG1可能是治疗OS的有效策略。版权所有©2023年作者。Elsevier B.V.出版。保留所有权利。
Dysregulation of long non-coding RNAs (lncRNAs) is an important component of tumorigenesis. Aberrant expression of lncRNA taurine upregulated gene 1 (lncTUG1) has been reported in various tumors; however, its precise role and key targets critically involved in osteosarcoma (OS) progression remains unclear.The expression profiles of lncRNAs and its regulated miRNAs related to OS progression were assessed by bioinformatics analysis and confirmed by qRT-PCR of OS cells. The miRNA targets were identified by transcriptome sequencing and verified by luciferase reporter and RNA pull-down assays. Several in vivo and in vitro approaches, including CCK8 assay, western blot, qRT-PCR, lentiviral transduction and OS cell xenograft mouse model were established to validate the effects of lncTUG1 regulation of miRNA and the downstream target genes on OS cell growth, apoptosis and progression.We found that lncTUG1 and miR-26a-5p were inversely up or down-regulated in OS cells, and siRNA-mediated lncTUG1 knockdown reversed the miR-26a-5p down-regulation and suppressed proliferation and enhanced apoptosis of OS cells. Further, we identified that an oncoprotein ZBTB7C was also upregulated in OS cells that were subjected to lncTUG1/miR-26a-5p regulation. More importantly, ZBTB7C knockdown reduced the ZBTB7C upregulation and ZBTB7C overexpression diminished the anti-OS effects of lncTUG1 knockdown in the OS xenograft model.Our data suggest that lncTUG1 acts as a miR-26a-5p sponge and promotes OS progression via up-regulating ZBTB7C, and targeting lncTUG1 might be an effective strategy to treat OS.Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.