Zhang Yaming, Hu Haili
Department of Otolaryngology, Huaihe Hospital of Henan University, Kaifeng, China.
Tumour Biol. 2017 Jun;39(6):1010428317699417. doi: 10.1177/1010428317699417.
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.
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。