Silencing of Long Non-coding RNA SMAD5-AS1 Reverses Epithelial Mesenchymal Transition in Nasopharyngeal Carcinoma via microRNA-195-Dependent Inhibition of SMAD5
Silencing of Long Non-coding RNA SMAD5-AS1 Reverses Epithelial Mesenchymal Transition in Nasopharyngeal Carcinoma via microRNA-195-Dependent Inhibition of SMAD5
复制标题
长非编码 RNA SMAD5-AS1 的沉默通过 microRNA-195 依赖性 SMAD5 抑制逆转鼻咽癌中的上皮间质转化
DOI：
10.3389/fonc.2019.01246
复制
发表时间：
2019-12-13
影响因子：
4.7
通讯作者：
Deng, Guangrui
中科院分区：
文献类型：
--
作者：
Li, Siwei;Zhao, Bo;Deng, Guangrui
近年来，长非编码RNA(LncRNAs)作为多种生物过程的关键调控因子得到了广泛的关注，但对其作用机制的了解仍然非常有限。以鼻咽癌和正常鼻咽癌组织中差异表达的lncRNA Smad5反义RNA1(Smad5-AS1)为研究对象，建议将microRNA-195(miR-195)与Smad5-AS1和Smad5结合。因此，本研究旨在探讨Smad5-AS1/miR-195/Smad5对鼻咽癌细胞上皮-间充质转化(EMT)的影响。RT-qPCR和Western印迹分析显示鼻咽癌组织和鼻咽癌细胞系中Smad5-AS1和Smad5高表达，miR-195低表达。Smad5-AS1通过与miR-195竞争性结合上调鼻咽癌细胞中Smad5的表达。随后在鼻咽癌细胞(CNE-2和CNE-1)中进行功能丧失和功能获得的研究，通过评估细胞生物学功能和体内致瘤能力以及检测EMT标志物的表达，探讨Smad5-AS、miR-195和Smad5在鼻咽癌进展中的作用。Smad5-AS1或Smad5下调或miR-195过表达可抑制鼻咽癌细胞的增殖、侵袭和迁移，逆转EMT，促进体外细胞凋亡，抑制体内肿瘤生长。总之，我们的研究结果表明，沉默lncRNA Smad5-AS1导致miR-195下调Smad5的表达，最终抑制鼻咽癌中的EMT。因此，Smad5-AS1可能是鼻咽癌干预治疗的潜在靶点。
Long non-coding RNAs (lncRNAs) have gained widespread attention in recent years as a key regulator of diverse biological processes, but the knowledge of the mechanisms by which they act is still very limited. Differentially expressed lncRNA SMAD5 antisense RNA 1 (SMAD5-AS1) in nasopharyngeal carcinoma (NPC) and normal samples shown by in silico analyses were selected as the main subject, and then microRNA-195 (miR-195) was suggested to bind to SMAD5-AS1 and SMAD5. Therefore, the purpose of the present study was to investigate the effects of SMAD5-AS1/miR-195/SMAD5 on epithelial-mesenchymal transition (EMT) in NPC cells. High expression of SMAD5-AS1 and SMAD5 but low miR-195 expression was determined in NPC tissues and NPC cell lines by RT-qPCR and western blot analysis. SMAD5-AS1 could upregulate SMAD5 expression by competitively binding to miR-195 in NPC cells. Loss- and gain-of-function investigations were subsequently conducted in NPC cells (CNE-2 and CNE-1) to explore the role of SMAD5-AS, miR-195 and SMAD5 in NPC progression by assessing cellular biological functions and tumorigenic ability in vivo as well as determining the expression of EMT markers. Downregulation of SMAD5-AS1 or SMAD5 or overexpression of miR-195 led to inhibited NPC cell proliferation, invasion and migration and reversed EMT, enhanced apoptosis in vitro as well as restrained tumor growth in vivo. In conclusion, our findings indicate that silencing of lncRNA SMAD5-AS1 induces the downregulation of SMAD5 by miR-195, eventually repressing EMT in NPC. Hence, SMAD5-AS1 may represent a potential therapeutic target for NPC intervention.