长链非编码 RNA XIST 的下调通过调节 miR-212-3p/CBLL1 轴抑制非小细胞肺癌细胞的增殖、迁移、侵袭和 EMT。
Downregulation of long non-coding RNA XIST inhibits cell proliferation, migration, invasion and EMT by regulating miR-212-3p/CBLL1 axis in non-small cell lung cancer cells.
机构信息
Department of Oncology, The Affiliated Hospital of Hubei University of Arts and Science, Xiangyang Central Hospital, Xiangyang, Hubei, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Oct;23(19):8391-8402. doi: 10.26355/eurrev_201910_19150.
OBJECTIVE
Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors in the world and its 5-year survival rate is very low. Long non-coding RNA X-inactive specific transcript (lncRNA XIST) has been demonstrated to play vital roles in NSCLC, but the exact molecular mechanisms underlying NSCLC still need to be further explored.
PATIENTS AND METHODS
Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) was performed to detect the expression of XIST, miR-212-3p and Casitas B-lineage proto-oncogene like 1 (CBLL1). Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were conducted to analyze the relationship among XIST, miR-212-3p and CBLL1. Cell Counting Kit-8 (CCK-8) assay and transwell invasion assay were carried out to evaluate cell proliferation, migration and invasion, respectively. Western blot analysis was conducted to examine the protein expression of CBLL1, E-cadherin, N-cadherin and Vimentin. Murine xenograft assay was conducted to explore the role of XIST in vivo.
RESULTS
Expression levels of XIST and CBLL1 were markedly upregulated, while the miR-212-3p level was markedly downregulated in NSCLC tissues and cells. MiR-212-3p was identified as a direct target of XIST, and miR-212-3p was predicted to target CBLL1. XIST knockdown repressed NSCLC cell proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) in vitro, and suppressed tumor growth in vivo, while miR-212-3p inhibition restored the effects. Furthermore, CBLL1 overexpression could abolish the effects of miR-212-3p overexpression on NSCLC cell proliferation, migration, invasion and EMT in vitro.
CONCLUSIONS
XIST was significantly decreased in NSCLC tissues and cells, and XIST knockdown suppressed the proliferation, migration, invasion and EMT of NSCLC cells by miR-212-3p/CBLL1 axis. These findings facilitated our understanding of lncRNA regulation in NSCLC.
目的
非小细胞肺癌（NSCLC）是世界上最常见的恶性肿瘤之一，其 5 年生存率非常低。长链非编码 RNA X 失活特异性转录物（lncRNA XIST）已被证明在 NSCLC 中发挥重要作用，但 NSCLC 的具体分子机制仍需进一步探索。
患者与方法
采用实时定量聚合酶链反应（qRT-PCR）检测 XIST、miR-212-3p 和 Casitas B 细胞系原癌基因样 1（CBLL1）的表达。通过双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）实验分析 XIST、miR-212-3p 与 CBLL1 之间的关系。通过细胞计数试剂盒（CCK-8）检测、Transwell 侵袭实验分别评估细胞增殖、迁移和侵袭能力。通过 Western blot 检测 CBLL1、E-钙黏蛋白、N-钙黏蛋白和波形蛋白的蛋白表达。通过建立小鼠异种移植瘤模型探讨 XIST 在体内的作用。
结果
XIST 和 CBLL1 的表达水平在 NSCLC 组织和细胞中显著上调，而 miR-212-3p 的水平显著下调。miR-212-3p 被鉴定为 XIST 的直接靶标，miR-212-3p 被预测靶向 CBLL1。XIST 敲低抑制 NSCLC 细胞在体外的增殖、迁移、侵袭和上皮间质转化（EMT），并抑制体内肿瘤生长，而 miR-212-3p 抑制恢复了这些作用。此外，CBLL1 的过表达可以消除 miR-212-3p 过表达对 NSCLC 细胞在体外增殖、迁移、侵袭和 EMT 的影响。
结论
XIST 在 NSCLC 组织和细胞中显著下调，XIST 敲低通过 miR-212-3p/CBLL1 轴抑制 NSCLC 细胞的增殖、迁移、侵袭和 EMT。这些发现有助于我们理解 lncRNA 在 NSCLC 中的调控作用。
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