长链非编码 RNA ZFAS1 通过靶向 miR-10a/SKA1 通路促进肾透明细胞癌的增殖和转移。
Long non-coding RNA ZFAS1 promotes proliferation and metastasis of clear cell renal cell carcinoma via targeting miR-10a/SKA1 pathway.
机构信息
Department of Pathophysiology, College of Basic Medical Science, China Medical University, Shenyang, People's Republic of China.
Department of Pathophysiology, College of Basic Medical Science, China Medical University, Shenyang, People's Republic of China; Department of Urology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang, People's Republic of China.
出版信息
Biomed Pharmacother. 2019 Mar;111:917-925. doi: 10.1016/j.biopha.2018.12.143. Epub 2019 Jan 7.
BACKGROUND
LncRNA ZFAS1 (ZNFX1 antisense RNA1) plays key roles in the occurrence and progression of various cancers, including colorectal cancer, glioma, lung cancer, gastric cancer, and so on. To date, relatively little is known about its potential role in development and/or progression of clear cell renal cell carcinoma (ccRCC).
METHODS
Expression level of ZFAS1 and miR-10a in 60 ccRCC and 20 adjacent non-tumor tissues were determined by using qRT-PCR. The effect of knockdown of ZFAS1 on cell proliferation, migration and invasion were measured by CCK-8 assay, transwell migration and invasion assay, respectively. The correlation of ZFAS1 and miR-10a was analyzed by bioinformatics DIANA TOOLS. Protein and mRNA expression of spindle and kinetochore-associated protein 1(SKA1) were determined by western blot and qRT-PCR analysis, respectively. Interactions between ZFAS1 and miR-10a were verified by luciferase reporter assay and RNA immunoprecipitation (RIP) assay, and interactions between miR-10a and SKA1 was verified by a luciferase reporter assay.
RESULTS
We observed that high-level expression of ZFAS1 is positively correlated with poor prognosis and shorter overall survival (OS) in patients with ccRCC. Knockdown of ZFAS1 significantly suppressed proliferation, migration and invasion of ccRCC cells. Furthermore, miR-10a was identified as a target of ZFAS1. Silencing miR-10a could attenuate the ability of ZFAS1 in promoting proliferation and metastasis of ccRCC. Subsequently, our studies validated that SKA1, as a key downstream target protein for miR-10a, is responsible for the biological role of ZFAS1. ZFAS1 positively regulated SKA1 expression via sponging miR-10a.
CONCLUSIONS
Taken together, our findings suggest that ZFAS1 promotes growth and metastasis of ccRCC via targeting miR-10a/SKA1 pathway, which may represent a novel therapeutic target or biomarker for ccRCC.
背景
长链非编码 RNA ZFAS1（锌指蛋白 X1 反义 RNA1）在包括结直肠癌、神经胶质瘤、肺癌、胃癌等在内的多种癌症的发生和发展中发挥着关键作用。迄今为止，人们对其在透明细胞肾细胞癌（ccRCC）发生和进展中的潜在作用知之甚少。
方法
通过 qRT-PCR 检测 60 例 ccRCC 组织和 20 例癌旁非肿瘤组织中 ZFAS1 和 miR-10a 的表达水平。通过 CCK-8 实验、transwell 迁移和侵袭实验分别测定 ZFAS1 敲低对细胞增殖、迁移和侵袭的影响。通过生物信息学 DIANA TOOLS 分析 ZFAS1 和 miR-10a 的相关性。通过 Western blot 和 qRT-PCR 分析分别检测纺锤体和动粒相关蛋白 1（SKA1）的蛋白和 mRNA 表达。通过荧光素酶报告基因实验和 RNA 免疫沉淀（RIP）实验验证 ZFAS1 与 miR-10a 之间的相互作用，通过荧光素酶报告基因实验验证 miR-10a 与 SKA1 之间的相互作用。
结果
我们观察到，ZFAS1 的高表达与 ccRCC 患者的不良预后和总生存期（OS）较短呈正相关。ZFAS1 敲低显著抑制 ccRCC 细胞的增殖、迁移和侵袭。此外，miR-10a 被鉴定为 ZFAS1 的靶标。沉默 miR-10a 可减弱 ZFAS1 促进 ccRCC 增殖和转移的能力。随后，我们的研究验证了 SKA1，作为 miR-10a 的关键下游靶蛋白，是 ZFAS1 发挥生物学作用的关键。ZFAS1 通过海绵 miR-10a 正向调节 SKA1 的表达。
结论
综上所述，我们的研究结果表明，ZFAS1 通过靶向 miR-10a/SKA1 通路促进 ccRCC 的生长和转移，这可能代表 ccRCC 的一种新的治疗靶点或生物标志物。
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