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长链非编码RNA CCAT1/miR-218/ZFX轴调控喉鳞状细胞癌的进展。

Long non-coding RNA CCAT1/miR-218/ZFX axis modulates the progression of laryngeal squamous cell cancer.

作者信息

Zhang Yaming, Hu Haili

机构信息

Department of Otolaryngology, Huaihe Hospital of Henan University, Kaifeng, China.

出版信息

Tumour Biol. 2017 Jun;39(6):1010428317699417. doi: 10.1177/1010428317699417.

Abstract

Long non-coding RNAs have been proved to be closely associated with different cancers. This study was designed to elucidate the function and mechanisms of colon cancer-associated transcript-1 in the progression of human laryngeal squamous cell cancer. Expressions of colon cancer-associated transcript-1, microRNA-218, and zinc finger protein, X-linked messenger RNA were measured using quantitative real-time polymerase chain reaction, and the expression level of zinc finger protein, X-linked protein was detected using western blot. Proliferation and invasion of laryngeal squamous cell cancer cell lines were detected by Cell Counting Kit-8 assay and Transwell invasion assay, respectively. Luciferase assay was used to confirm whether microRNA-218 is a target of colon cancer-associated transcript-1 and whether microRNA-218 directly binds to 3'-untranslated region of zinc finger protein, X-linked messenger RNA. Effect of colon cancer-associated transcript-1 on tumor growth was observed through xenograft mice models in vivo. The results showed that expressions of colon cancer-associated transcript-1 and zinc finger protein, X-linked were significantly higher while microRNA-218 expression was significantly lower in the laryngeal squamous cell cancer tissues than those in the adjacent normal tissues. MicroRNA-218 overexpression or zinc finger protein, X-linked silencing significantly suppressed proliferation and invasion of laryngeal squamous cell cancer cells. Moreover, knockdown of colon cancer-associated transcript-1 significantly inhibited proliferation and invasion of laryngeal squamous cell cancer cells, which were reversed by microRNA-218 downregulation or zinc finger protein, X-linked upregulation. Finally, colon cancer-associated transcript-1 silencing inhibited xenograft tumor growth of laryngeal squamous cell cancer in vivo. In conclusion, colon cancer-associated transcript-1 knockdown inhibits proliferation and invasion of laryngeal squamous cell cancer cells through enhancing zinc finger protein, X-linked by sponging microRNA-218, elucidating a novel colon cancer-associated transcript-1-microRNA-218-zinc finger protein, X-linked regulatory axis in laryngeal squamous cell cancer and providing a promising therapeutic target for laryngeal squamous cell cancer patients.

摘要

长链非编码RNA已被证明与不同癌症密切相关。本研究旨在阐明结肠癌相关转录本-1在人喉鳞状细胞癌进展中的功能及机制。采用定量实时聚合酶链反应检测结肠癌相关转录本-1、微小RNA-218和X连锁锌指蛋白信使核糖核酸的表达,并通过蛋白质免疫印迹法检测X连锁锌指蛋白的表达水平。分别采用细胞计数试剂盒-8法和Transwell侵袭实验检测喉鳞状细胞癌细胞系的增殖和侵袭能力。采用荧光素酶报告基因实验证实微小RNA-218是否为结肠癌相关转录本-1的靶标,以及微小RNA-218是否直接结合X连锁锌指蛋白信使核糖核酸的3'-非翻译区。通过体内异种移植小鼠模型观察结肠癌相关转录本-1对肿瘤生长的影响。结果显示,喉鳞状细胞癌组织中结肠癌相关转录本-1和X连锁锌指蛋白的表达明显高于相邻正常组织,而微小RNA-218的表达明显低于相邻正常组织。微小RNA-218过表达或X连锁锌指蛋白沉默显著抑制喉鳞状细胞癌细胞的增殖和侵袭。此外,敲低结肠癌相关转录本-1显著抑制喉鳞状细胞癌细胞的增殖和侵袭,而微小RNA-218下调或X连锁锌指蛋白上调可逆转这种抑制作用。最后,结肠癌相关转录本-1沉默抑制了喉鳞状细胞癌在体内的异种移植肿瘤生长。总之,敲低结肠癌相关转录本-1通过海绵吸附微小RNA-218增强X连锁锌指蛋白的表达,从而抑制喉鳞状细胞癌细胞的增殖和侵袭,阐明了喉鳞状细胞癌中一种新的结肠癌相关转录本-1-微小RNA-218-X连锁锌指蛋白调控轴,并为喉鳞状细胞癌患者提供了一个有前景的治疗靶点。

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