Background

Long non-coding RNAs (lncRNAs) are transcribed pervasively in the genome and act to regulate chromatin remodeling and gene expression. Dysregulated lncRNA expression has been reported in many cancers, but the role of lncRNAs in esophageal cancer (EC) has so far remained poorly understood. In this study, we aimed to understand the effect of lncRNA LINC01234 on EC development through competitively binding to microRNA-193a-5p (miR-193a-5p).

Methods

The Gene Expression Omnibus (GEO) database was used for microarray-based EC expression profiling. Gain- and loss-of-function analyses were carried out in human EC-derived Eca-109 and EC9706 cells. Expression analyses of miR-193a-5p, LINC01234, CCNE1, caspase-3, p21, Bax, cyclinD1 and Bcl-2 were performed using RT-qPCR and Western blotting. Cell proliferation, colony formation and apoptosis analyses were carried out using MTT, Hoechst 33258 and flow cytometry assays. A xenograft EC model in nude mice was used to evaluate in vivo tumor growth and CCNE1 expression.

Results

Microarray-based analyses revealed that LINC01234 expression was increased in primary EC samples, whereas that of miR-193a-5p was decreased. We found that CCNE1 was a target of miR-193a-5p and that LINC01234, in turn, sponges miR-193a-5p. After treatment with si-LINC01234 or miR-193a-5p mimic, EC cells (Eca-109 and EC9706) exhibited cyclinD1 and Bcl-2 downregulation, and caspase-3, p21, Bax and cleaved caspase-3 upregulation. LINC01234 silencing or miR-193a-5p upregulation resulted in decreased proliferation and colony formation, and increased apoptosis of EC cells. In addition, LINC01234 silencing or miR-193a-5p upregulation resulted in reduced in vivo EC tumor growth and CCNE1 expression in nude mice.

Conclusions

We found that silencing of LINC01234 suppresses EC development by inhibiting CCNE1 through competitively binding to miR-193a-5p, which suggests that LINC01234 may represent a novel target for EC therapy.

中文翻译：

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长时间的非编码RNA LINC01234沉默通过microRNA-193a-5p介导的CCNE1下调在食道癌细胞中发挥抗癌作用。

背景

长的非编码RNA（lncRNA）在基因组中普遍转录，并起着调节染色质重塑和基因表达的作用。已经在许多癌症中报道了lncRNA表达失调，但是迄今为止，对nccRNA在食道癌（EC）中的作用还知之甚少。在这项研究中，我们旨在通过竞争性结合microRNA-193a-5p（miR-193a-5p）来了解lncRNA LINC01234对EC发育的影响。

方法

基因表达综合（GEO）数据库用于基于微阵列的EC表达谱分析。在源自人EC的Eca-109和EC9706细胞中进行了功能获得和功能丧失分析。使用RT-qPCR和Western印迹进行miR-193a-5p，LINC01234，CCNE1，caspase-3，p21，Bax，cyclinD1和Bcl-2的表达分析。使用MTT，Hoechst 33258和流式细胞仪进行细胞增殖，集落形成和凋亡分析。裸鼠的异种移植EC模型用于评估体内肿瘤生长和CCNE1表达。

结果

基于微阵列的分析表明，LINC01234表达在原代EC样品中增加，而miR-193a-5p则下降。我们发现CCNE1是miR-193a-5p的靶标，而LINC01234反过来是miR-193a-5p的海绵。用si-LINC01234或miR-193a-5p模拟物处理后，EC细胞（Eca-109和EC9706）表现出cyclinD1和Bcl-2下调，而caspase-3，p21，Bax和裂解的caspase-3上调。LINC01234沉默或miR-193a-5p上调导致减少的增殖和集落形成，并增加EC细胞的凋亡。此外，LINC01234沉默或miR-193a-5p上调导致裸鼠体内EC肿瘤生长和CCNE1表达降低。

结论

我们发现沉默LINC01234可通过竞争结合miR-193a-5p抑制CCNE1来抑制EC的发展，这表明LINC01234可能代表EC治疗的新靶点。