Long non-coding RNA NKILA inhibits migration and invasion of non-small cell lung cancer via NF-κB/Snail pathway.
Long non-coding RNA NKILA inhibits migration and invasion of non-small cell lung cancer via NF-κB/Snail pathway.
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DOI：
10.1186/s13046-017-0518-0
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发表时间：
2017-04-17
期刊：
影响因子：
--
通讯作者：
He J
中科院分区：
文献类型：
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作者：
Lu Z;Li Y;Wang J;Che Y;Sun S;Huang J;Chen Z;He J
大量研究表明，长链非编码RNA（longnoncodingRNAs，lncRNA）在肿瘤细胞增殖、凋亡、迁移和侵袭等过程中发挥着重要作用。以往的研究发现NKILA与NF-κ B的核转位相互作用并抑制NF-κ B的核转位，从而影响乳腺癌的转移和预后。然而，NKILA在非小细胞肺癌（NSCLC）中的临床意义和生物学作用仍然未知。我们检测了106对NSCLC组织和细胞系中NKILA的表达水平。还通过qRT-PCR检测TGF-β1刺激后NKILA的表达水平，并通过染色质免疫沉淀（ChIP）进行验证。进行功能获得和功能丧失测定以检查NKILA对NSCLC细胞增殖、迁移和侵袭的影响。通过RNA免疫沉淀（RIP）、蛋白质印迹（western blot）和拯救实验，研究NKILA、NF-κB与EMT信号通路的关系。NKILA在NSCLC组织中的表达明显低于癌旁组织，且NKILA的低表达与淋巴结转移和TNM分期密切相关。我们发现NKILA在NSCLC细胞中的表达主要受经典的TGF-β信号通路的调控，而不是乳腺癌中报道的NF-κB通路。功能获得和丧失测定发现NKILA抑制NSCLC细胞的迁移、侵袭和活力。NKILA通过抑制IκBα的磷酸化和NF-κB的活化，抑制Snail的表达，进而抑制上皮-间质转化过程标志物的表达。本研究发现NKILA在非小细胞肺癌组织中表达下调，促进了非小细胞肺癌的转移。体外研究进一步阐明NKILA的表达通过经典的TGF-β信号通路调节，进而通过干扰NF-κB/Snail信号通路抑制NSCLC细胞的迁移和侵袭。本文的在线版本（doi：10.1186/s13046-017-0518-0）包含补充材料，可供授权用户使用。
Numerous studies have shown that long non-coding RNAs (lncRNAs) play key roles during multiple cancer processes, such as cell proliferation, apoptosis, migration and invasion. The previous studies found that NKILA interacted with and suppressed the nuclear translocation of NF-KappaB, which influenced metastasis and prognosis in breast cancer. However the clinical significance and biological role of NKILA in non-small cell lung cancer (NSCLC) remains unknown. We examined expression levels of NKILA in 106 pairs of NSCLC tissues and cell lines. The expression level of NKILA after TGF-β1 stimulation also was examined by qRT-PCR and validated by Chromatin immunoprecipitation (ChIP). Gain-of-function and loss-of-function assays were performed to examine the effect of NKILA on proliferation, migration and invasion of NSCLC cells. RNA immunoprecipitation (RIP), western blot and rescue experiments were carried out to reveal the interrelation between NKILA, NF-κB and EMT signal pathway. The expression of NKILA was down-regulated in NSCLC cancer tissues compared with matched adjacent noncancerous tissues, and lower NKILA expression in tumor tissues were significantly correlated with lymph node metastasis and advanced TNM stage. We found that the expression of NKILA was mainly regulated by classical TGF-β signal pathway in NSCLC cells rather than NF-κB pathway reported in breast cancer. Gain and loss of function assays found that NKILA inhibited migration, invasion and viability of NSCLC cells. Mechanistic study showed that NKILA attenuated Snail expression via inhibiting the phosphorylation of IκBα and NF-κB activation, subsequently suppressed the expression of markers of epithelial-mesenchymal transition process. The present study found that the expression of NKILA was downregulated in tumor tissues of NSCLC, which improved the metastasis of NSCLC patients. In vitro studies further clarified that the expression of NKILA was regulated through classical TGF-β signal pathway, which subsequently inhibited migration and invasion of NSCLC cells through interfering NF-κB/Snail signal pathway in NSCLC cells. The online version of this article (doi:10.1186/s13046-017-0518-0) contains supplementary material, which is available to authorized users.