长链非编码 RNA HOTAIR 通过海绵吸附 microRNA-124 促进α-2,8-唾液酸转移酶 4 促进肾细胞癌恶性转化。
Long noncoding RNA HOTAIR promotes renal cell carcinoma malignancy through alpha-2, 8-sialyltransferase 4 by sponging microRNA-124.
机构信息
College of Laboratory Medicine, Dalian Medical University, Dalian, Liaoning Province, China.
Benxi Jinshan Hospital, Benxi, Liaoning Province, China.
出版信息
Cell Prolif. 2018 Dec;51(6):e12507. doi: 10.1111/cpr.12507. Epub 2018 Aug 13.
OBJECTIVES
Accumulating evidence demonstrated that the long noncoding RNA (lncRNA) HOTAIR (Hox transcript antisense intergenic RNA) plays key role in renal cell carcinoma (RCC) malignancy, while microRNA-124 (miR-124) is a tumour suppressor in RCC. The aim of this work was to assess the biological function of HOTAIR and to explore underlying mechanism involved in HOTAIR/miR-124/alpha-2, 8-sialyltransferase 4 (ST8SIA4) axis-regulated progression in RCC.
MATERIALS AND METHODS
Real-time PCR analyses and western blots were performed to the levels of HOTAIR, miR-124 and ST8SIA4 expression in human RCC tissues and RCC cell lines (ACHN and 786-O). Bioinformatics analysis and dual-luciferase reporter assay were used to illustrate relationship between HOTAIR and miR-124 in RCC. Colony formation assays, EdU assays, Ki67 assays and apoptosis assays were taken to evaluate cell proliferation. Tumour xenograft was created to explore the functions of HOTAIR and ST8SIA4 in tumorigenesis in vivo. Migration assays, invasion assays and cell adhesion assays and were also taken to analyse the carcinoma progression.
RESULTS
In this study, HOTAIR level was confirmed to be significantly upregulated in RCC samples and RCC cell lines compared with those in the paired adjacent tissues and normal renal cell line. Overexpression of HOTAIR promoted the capability of proliferation, migration and invasion in RCC cell lines. HOTAIR directly bound to miR-124, while miR-124 mediated the expression of ST8SIA4 in RCC cell lines. ST8SIA4 was upregulated in RCC tissues and RCC cell lines. Ectopic expression of ST8SIA4 modulated the proliferation, migration and invasion of RCC cells. Further results indicated that HOTAIR promoted the proliferation and metastasis as a competing endogenous RNA to regulate ST8SIA4 expression by sponging miR-124 in RCC.
CONCLUSIONS
Our results demonstrated that HOTAIR mediated RCC progression in part through miR-124/ST8SIA4 axis, which functioned as a new prognostic biomarker in RCC.
目的
越来越多的证据表明，长链非编码 RNA（lncRNA）HOTAIR（HOX 转录反义基因间 RNA）在肾细胞癌（RCC）恶性肿瘤中发挥关键作用，而 microRNA-124（miR-124）是 RCC 的肿瘤抑制因子。本研究旨在评估 HOTAIR 的生物学功能，并探讨 HOTAIR/miR-124/α-2,8-唾液酸转移酶 4（ST8SIA4）轴调控 RCC 进展的潜在机制。
材料和方法
采用实时 PCR 分析和 Western blot 检测人 RCC 组织和 RCC 细胞系（ACHN 和 786-O）中 HOTAIR、miR-124 和 ST8SIA4 的表达水平。生物信息学分析和双荧光素酶报告基因实验用于阐明 RCC 中 HOTAIR 和 miR-124 之间的关系。集落形成实验、EdU 实验、Ki67 实验和凋亡实验用于评估细胞增殖。建立肿瘤异种移植模型以研究 HOTAIR 和 ST8SIA4 在体内肿瘤发生中的作用。迁移实验、侵袭实验和细胞黏附实验用于分析癌细胞的进展。
结果
本研究证实，与配对的相邻组织和正常肾细胞系相比，HOTAIR 在 RCC 样本和 RCC 细胞系中的表达水平显著上调。HOTAIR 的过表达促进了 RCC 细胞系的增殖、迁移和侵袭能力。HOTAIR 可直接与 miR-124 结合，而 miR-124 在 RCC 细胞系中介导 ST8SIA4 的表达。ST8SIA4 在 RCC 组织和 RCC 细胞系中上调。外源性表达 ST8SIA4 可调节 RCC 细胞的增殖、迁移和侵袭。进一步的结果表明，HOTAIR 通过作为竞争性内源性 RNA 来调节 ST8SIA4 的表达，从而促进 RCC 的增殖和转移，通过海绵吸附 miR-124。
结论
我们的研究结果表明，HOTAIR 通过 miR-124/ST8SIA4 轴介导 RCC 进展，可作为 RCC 的一种新的预后生物标志物。
Zotero 插件