Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, USA.
Nucleic Acids Res. 2013 May;41(9):4976-87. doi: 10.1093/nar/gkt182. Epub 2013 Apr 4.
Protein-coding genes account for only a small part of the human genome, whereas the vast majority of transcripts make up the non-coding RNAs including long non-coding RNAs (lncRNAs). Accumulating evidence indicates that lncRNAs could play a critical role in regulation of cellular processes such as cell growth and apoptosis as well as cancer progression and metastasis. LncRNA loc285194 was previously shown to be within a tumor suppressor unit in osteosarcoma and to suppress tumor cell growth. However, it is unknown regarding the regulation of loc285194. Moreover, the underlying mechanism by which loc285194 functions as a potential tumor suppressor is elusive. In this study, we show that loc285194 is a p53 transcription target; ectopic expression of loc285194 inhibits tumor cell growth both in vitro and in vivo. Through deletion analysis, we identify an active region responsible for tumor cell growth inhibition within exon 4, which harbors two miR-211 binding sites. Importantly, this loc285194-mediated growth inhibition is in part due to specific suppression of miR-211. We further demonstrate a reciprocal repression between loc285194 and miR-211; in contrast to loc285194, miR-211 promotes cell growth. Finally, we detect downregulation of loc285194 in colon cancer specimens by quantitative PCR arrays and in situ hybridization of tissue microarrays. Together, these results suggest that loc285194 is a p53-regulated tumor suppressor, which acts in part through repression of miR-211.
蛋白编码基因仅占人类基因组的一小部分,而绝大多数转录本构成非编码 RNA,包括长非编码 RNA(lncRNA)。越来越多的证据表明,lncRNA 可以在细胞过程的调节中发挥关键作用,如细胞生长和凋亡以及癌症的进展和转移。先前已经表明 lncRNA loc285194 位于骨肉瘤中的肿瘤抑制单元内,并抑制肿瘤细胞生长。然而,关于 loc285194 的调节尚不清楚。此外,loc285194 作为潜在的肿瘤抑制因子的作用机制尚不清楚。在这项研究中,我们表明 loc285194 是 p53 转录靶标;loc285194 的异位表达在体外和体内均抑制肿瘤细胞生长。通过缺失分析,我们确定了在exon4 内负责肿瘤细胞生长抑制的活性区域,该区域包含两个 miR-211 结合位点。重要的是,这种 loc285194 介导的生长抑制部分归因于对 miR-211 的特异性抑制。我们进一步证明了 loc285194 和 miR-211 之间的相互抑制;与 loc285194 相反,miR-211 促进细胞生长。最后,我们通过定量 PCR 阵列和组织微阵列的原位杂交检测到结肠癌标本中 loc285194 的下调。总之,这些结果表明 loc285194 是一种 p53 调节的肿瘤抑制因子,部分通过抑制 miR-211 起作用。