Song Hu, Xu Yixin, Shi Linseng, Xu Teng, Fan Ruizhi, Cao Meng, Xu Wei, Song Jun
Department of General Surgery, the Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou, Jiangsu, 221002, PR China; Institute of Digestive Disease, Xuzhou Medical University, 84 West Huaihai Road, Xuzhou, Jiangsu, 221002, PR China.
Department of General Surgery, the Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou, Jiangsu, 221002, PR China; Institute of Digestive Disease, Xuzhou Medical University, 84 West Huaihai Road, Xuzhou, Jiangsu, 221002, PR China.
Biomed Pharmacother. 2018 Dec;108:338-346. doi: 10.1016/j.biopha.2018.09.057. Epub 2018 Sep 15.
This work aims to explore the roles and mechanisms of long non coding RNA (lncRNA) THOR in regulating the stemness of gastric cancer cells. RNA-sequencing combined with quantitative real-time PCR (qRT-PCR) indicated that lncRNA THOR level was significantly upregulated in gastric cancer tissues compared with that in normal adjacent tissues. Knockdown of THOR attenuated the stemnness of gastric cancer cells, evident by the decrease of stemness markers expression and capacity of cells spheroid formation. Further RNA-sequencing combined with qRT-PCR and western blot analysis demonstrated that expression of transcriptional factor SOX9 was remarkably decreased in gastric cancer cells with THOR stable knockdown. Additionally, RNA immunoprecipitation (RIP) combined with luciferase reporter assay revealed that THOR directly bound to SOX9 3' untranslated region (3'UTR), but not its 5'UTR or coding area. Notably, overexpression of SOX9 rescued THOR knockdown-mediated inhibition on the stemness of gastric cancer cells. Thus, our results suggest that THOR could potentiate the stemness of gastric cancer cells via directly binding to SOX9 3'UTR.
这项研究旨在探索长链非编码RNA（lncRNA）THOR在调控胃癌细胞干性中的作用及机制。RNA测序结合定量实时聚合酶链反应（qRT-PCR）表明，与癌旁正常组织相比，lncRNA THOR在胃癌组织中的表达水平显著上调。敲低THOR可减弱胃癌细胞的干性，这表现为干性标志物表达降低以及细胞球形成能力下降。进一步的RNA测序结合qRT-PCR和蛋白质免疫印迹分析表明，在THOR稳定敲低的胃癌细胞中，转录因子SOX9的表达显著降低。此外，RNA免疫沉淀（RIP）结合荧光素酶报告基因检测显示，THOR直接与SOX9的3'非翻译区（3'UTR）结合，而非其5'UTR或编码区。值得注意的是，SOX9的过表达挽救了THOR敲低介导的对胃癌细胞干性的抑制作用。因此，我们的研究结果表明，THOR可通过直接结合SOX9的3'UTR增强胃癌细胞的干性。
