BACKGROUND The CDKN2A/B locus contains crucial tumor suppressors and a lncRNA gene ANRIL. However, the mechanisms that coordinately regulate their expression levels are not clear. METHODS Novel RNAs transcribed from the CDKN2A gene were screened by CDKN2A-specific RNA capture deep-sequencing and confirmed by Northern blotting and clone-sequencing. Long non-coding RNA (lncRNA) binding proteins were characterized by RNA pull-down combined with mass spectrometry and RNA immunoprecipitation. LncRNA functions in human cells were studied using a set of biological assays in vitro and in vivo. RESULTS We characterized a novel lncRNA, P14AS with its promoter in the antisense strand of the fragment near CDKN2A exon 1b in human cells. The mature P14AS is a three-exon linear cytoplasmic lncRNA (1043-nt), including an AU-rich element (ARE) in exon 1. P14AS decreases AUF1-ANRIL/P16 RNA interaction and then increases ANRIL/P16 expression by competitively binding to AUF1 P37 and P40 isoforms. Interestingly, P14AS significantly promoted the proliferation of cancer cells and tumor formation in NOD-SCID mice in a P16-independent pattern. Moreover, in human colon cancer tissues, the expression levels of P14AS and ANRIL lncRNAs were significantly upregulated compared with the paired normal tissues. CONCLUSION A novel lncRNA, P14AS, transcribed from the antisense strand of the CDKN2A/P14 gene, promotes colon cancer development by cis upregulating the expression of oncogenic ANRIL.

中文翻译：

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通过与人细胞中的AUF1结合来表征新型LncRNA P14AS作为ANRIL的保护剂。

背景技术CDKN2A / B基因座包含关键的肿瘤抑制子和lncRNA基因ANRIL。但是，协调调节其表达水平的机制尚不清楚。方法通过CDKN2A特异性RNA捕获深度测序筛选从CDKN2A基因转录得到的新型RNA，并通过Northern印迹和克隆测序进行确认。长非编码RNA（lncRNA）结合蛋白的特征是RNA下拉结合质谱和RNA免疫沉淀。在体外和体内使用一组生物学测定法研究了人类细胞中的LncRNA功能。结果我们表征了一种新型的lncRNA P14AS，其启动子位于人类细胞中CDKN2A外显子1b附近片段的反义链中。成熟的P14AS是三外显子线性胞质lncRNA（1043-nt），在外显子1中包含富含AU的元件（ARE）。P14AS通过竞争性结合AUF1 P37和P40同工型降低AUF1-ANRIL / P16 RNA相互作用，然后增加ANRIL / P16表达。有趣的是，P14AS以非P16的方式显着促进了NOD-SCID小鼠中癌细胞的增殖和肿瘤的形成。此外，在人类结肠癌组织中，与配对的正常组织相比，P14AS和ANRIL lncRNA的表达水平显着上调。结论从CDKN2A / P14基因反义链转录的新型lncRNA P14AS通过顺式上调致癌性ANRIL的表达促进结肠癌的发展。P14AS以非P16依赖性模式显着促进NOD-SCID小鼠体内癌细胞的增殖和肿瘤形成。此外，在人类结肠癌组织中，与配对的正常组织相比，P14AS和ANRIL lncRNA的表达水平显着上调。结论从CDKN2A / P14基因反义链转录的新型lncRNA P14AS通过顺式上调致癌性ANRIL的表达促进结肠癌的发展。P14AS以非P16方式显着促进NOD-SCID小鼠中癌细胞的增殖和肿瘤形成。此外，在人类结肠癌组织中，与配对的正常组织相比，P14AS和ANRIL lncRNA的表达水平显着上调。结论从CDKN2A / P14基因反义链转录的新型lncRNA P14AS通过顺式上调致癌性ANRIL的表达促进结肠癌的发展。