Liu Guanghua, Ye Zixing, Zhao Xin, Ji Zhigang
Department of Urology, Peking Union Medical College Hospital, Chinese Academy of Medical ScienceBeijing 100730, China.
Am J Cancer Res. 2017 Dec 1;7(12):2515-2525. eCollection 2017.
Clear cell renal cell carcinoma (ccRCC) is one of the leading causes of genitourinary cancer-related death, largely due to the metastasis of ccRCC. Previous profiling study showed that lncRNAs are critical regulators in lots of cancers. However, the roles of specific lncRNAs in ccRCC migration and invasion are still unknown. In this study, we utilized the high-throughput genome sequencing to identify the potential differentially expressed lncRNAs in ccRCC and further determined the underlying regulatory mechanism. We found that lncRNA SNHG14 was significantly up-regulated in ccRCC cell lines in contrast to normal renal epithelial cells. By performing bioinformatics analysis and luciferase reporter assays, we revealed that the transcription factor SP1 can bind to the promoter region of SNHG14, resulting in the overexpression of SNHG14 in ccRCC. Functionally, enhanced expression of lncRNA SNHG14 promoted cell migration and invasion through promoting N-WASP protein level. More importantly, RT-qPCR and in situ RNA FISH analysis showed that SNHG14 was predominantly abundant in the cytoplasm of ccRCC cells. The subsequent RNA immunoprecipitation assay, and gain or loss-function assays showed that SNHG14 functioned as ceRNA to regulate N-WASP expression and cell motility ability via a miR-203-dependent manner. Our results imply that SNHG14 is a critical lncRNA that promotes ccRCC migration and invasion via sponging miR-203 and elevating N-WASP. Therefore, SNHG14 could serve as a promising therapeutic target for ccRCC.
透明细胞肾细胞癌（ccRCC）是泌尿生殖系统癌症相关死亡的主要原因之一，这主要归因于ccRCC的转移。先前的分析研究表明，长链非编码RNA（lncRNAs）是许多癌症中的关键调节因子。然而，特定lncRNAs在ccRCC迁移和侵袭中的作用仍然未知。在本研究中，我们利用高通量基因组测序来鉴定ccRCC中潜在的差异表达lncRNAs，并进一步确定其潜在的调控机制。我们发现，与正常肾上皮细胞相比，lncRNA SNHG14在ccRCC细胞系中显著上调。通过进行生物信息学分析和荧光素酶报告基因检测，我们揭示转录因子SP1可以结合SNHG14的启动子区域，导致SNHG14在ccRCC中过表达。在功能上，lncRNA SNHG14的表达增强通过促进N-WASP蛋白水平来促进细胞迁移和侵袭。更重要的是，RT-qPCR和原位RNA FISH分析表明，SNHG14在ccRCC细胞的细胞质中主要大量存在。随后的RNA免疫沉淀试验以及功能获得或丧失试验表明，SNHG14作为竞争性内源RNA（ceRNA）通过miR-203依赖的方式调节N-WASP表达和细胞运动能力。我们的结果表明，SNHG14是一种关键的lncRNA，通过结合miR-203并提高N-WASP水平来促进ccRCC的迁移和侵袭。因此，SNHG14有望成为ccRCC的治疗靶点。
