Abstract：
BACKGROUND: Long non-coding RNAs (lncRNAs) are increasingly observed as regulatory factors for the initiation and progression of varying kinds of cancers. However, studies on lncRNAs in non-small cell lung cancer (NSCLC) progression are currently lacking.
OBJECTIVE: We intended to determine the role of lncRNA LINC00472 and its downstream regulatory mechanism in NSCLC, thus providing novel ideas for targeted therapies for NSCLC.
METHODS: The target signaling axis comprising the lncRNA/microRNA/mRNA was identified through bioinformatics analysis. Subcellular localization of LINC00472 was assessed with fluorescence in situ hybridization (FISH). Cellular function experiments were conducted to examine the proliferation, migration, invasion, and apoptosis of NSCLC cells, and dual-luciferase and RNA binding protein immunoprecipitation assays were performed to validate the binding relationship. Quantitative real-time polymerase chain reaction (qPCR) and western blot were utilized to assess the expression levels of the investigated gene and protein, respectively.
RESULTS: The LINC00472 expression was markedly decreased in NSCLC tissues and cells. The FISH, combined with nuclear-cytoplasm separation assay, demonstrated that LINC00472 was mainly located in the cytoplasm. The overexpression of LINC00472 restrained proliferation and metastasis of NSCLC in vitro. The LINC00472 could target and repress miR-1275 level, and overexpression of LINC00472 reduced the miR-1275-dependent malignant cell phenotype in NSCLC. Further study revealed that HOXA2 was a downstream target of miR-1275 and was negatively modulated by miR-1275. Rescue assays exhibited that the overexpression of miR-1275 or inhibition of HOXA2 reversed the impact of LINC00472 overexpression on the malignant progression of NSCLC cells. The LINC00472 repressed the epithelial-mesenchymal transition (EMT) of NSCLC cells through miR-1275/HOXA2.
CONCLUSIONS: The LINC00472 functioned as a competing endogenous RNA to modulate HOXA2 level by sponging miR-1275 in NSCLC. Simultaneously, the LINC00472/miR-1275/HOXA2 axis may be a possible therapeutic target and biomarker for NSCLC.
OBJECTIVE: We intended to determine the role of lncRNA LINC00472 and its downstream regulatory mechanism in NSCLC, thus providing novel ideas for targeted therapies for NSCLC.
METHODS: The target signaling axis comprising the lncRNA/microRNA/mRNA was identified through bioinformatics analysis. Subcellular localization of LINC00472 was assessed with fluorescence in situ hybridization (FISH). Cellular function experiments were conducted to examine the proliferation, migration, invasion, and apoptosis of NSCLC cells, and dual-luciferase and RNA binding protein immunoprecipitation assays were performed to validate the binding relationship. Quantitative real-time polymerase chain reaction (qPCR) and western blot were utilized to assess the expression levels of the investigated gene and protein, respectively.
RESULTS: The LINC00472 expression was markedly decreased in NSCLC tissues and cells. The FISH, combined with nuclear-cytoplasm separation assay, demonstrated that LINC00472 was mainly located in the cytoplasm. The overexpression of LINC00472 restrained proliferation and metastasis of NSCLC in vitro. The LINC00472 could target and repress miR-1275 level, and overexpression of LINC00472 reduced the miR-1275-dependent malignant cell phenotype in NSCLC. Further study revealed that HOXA2 was a downstream target of miR-1275 and was negatively modulated by miR-1275. Rescue assays exhibited that the overexpression of miR-1275 or inhibition of HOXA2 reversed the impact of LINC00472 overexpression on the malignant progression of NSCLC cells. The LINC00472 repressed the epithelial-mesenchymal transition (EMT) of NSCLC cells through miR-1275/HOXA2.
CONCLUSIONS: The LINC00472 functioned as a competing endogenous RNA to modulate HOXA2 level by sponging miR-1275 in NSCLC. Simultaneously, the LINC00472/miR-1275/HOXA2 axis may be a possible therapeutic target and biomarker for NSCLC.
摘要：
背景：长链非编码RNA（lncRNA）越来越多地被观察到作为不同类型癌症的起始和进展的调节因子。然而,目前缺乏lncRNAs在非小细胞肺癌(NSCLC)进展中的研究.
目的：我们打算确定lncRNALINC00472及其下游调控机制在NSCLC中的作用，从而为NSCLC的靶向治疗提供新思路。
方法：通过生物信息学分析鉴定了包含lncRNA/microRNA/mRNA的靶信号轴。用荧光原位杂交(FISH)评估LINC00472的亚细胞定位。进行细胞功能实验以检查增殖，迁移,入侵,和NSCLC细胞凋亡，并进行双荧光素酶和RNA结合蛋白免疫沉淀测定以验证结合关系。定量实时聚合酶链反应（qPCR）和蛋白质印迹用于评估研究的基因和蛋白质的表达水平，分别。
结果：LINC00472在NSCLC组织和细胞中的表达显著降低。鱼,结合核质分离试验，证明LINC00472主要位于细胞质中。LINC00472的过表达在体外克制NSCLC的增殖和转移。LINC00472可以靶向和抑制miR-1275水平，LINC00472的过表达降低了NSCLC中miR-1275依赖性恶性细胞表型。进一步的研究表明，HOXA2是miR-1275的下游靶标，并被miR-1275负调节。挽救测定显示miR-1275的过表达或HOXA2的抑制逆转了LINC00472过表达对NSCLC细胞的恶性进展的影响。LINC00472通过miR-1275/HOXA2抑制NSCLC细胞的上皮-间质转化(EMT)。
结论：LINC00472作为竞争性内源性RNA，通过在NSCLC中形成miR-1275来调节HOXA2水平。同时,LINC00472/miR-1275/HOXA2轴可能是NSCLC的治疗靶点和生物标志物.
目的：我们打算确定lncRNALINC00472及其下游调控机制在NSCLC中的作用，从而为NSCLC的靶向治疗提供新思路。
方法：通过生物信息学分析鉴定了包含lncRNA/microRNA/mRNA的靶信号轴。用荧光原位杂交(FISH)评估LINC00472的亚细胞定位。进行细胞功能实验以检查增殖，迁移,入侵,和NSCLC细胞凋亡，并进行双荧光素酶和RNA结合蛋白免疫沉淀测定以验证结合关系。定量实时聚合酶链反应（qPCR）和蛋白质印迹用于评估研究的基因和蛋白质的表达水平，分别。
结果：LINC00472在NSCLC组织和细胞中的表达显著降低。鱼,结合核质分离试验，证明LINC00472主要位于细胞质中。LINC00472的过表达在体外克制NSCLC的增殖和转移。LINC00472可以靶向和抑制miR-1275水平，LINC00472的过表达降低了NSCLC中miR-1275依赖性恶性细胞表型。进一步的研究表明，HOXA2是miR-1275的下游靶标，并被miR-1275负调节。挽救测定显示miR-1275的过表达或HOXA2的抑制逆转了LINC00472过表达对NSCLC细胞的恶性进展的影响。LINC00472通过miR-1275/HOXA2抑制NSCLC细胞的上皮-间质转化(EMT)。
结论：LINC00472作为竞争性内源性RNA，通过在NSCLC中形成miR-1275来调节HOXA2水平。同时,LINC00472/miR-1275/HOXA2轴可能是NSCLC的治疗靶点和生物标志物.
