Wang Guanghui, Wang Shujing, Li Congying
1 Department of Otorhinolaryngology, Huaihe Hospital of Henan University, Kaifeng, China.
2 Department of Otorhinolaryngology, School of Medicine, Kaifeng University, Kaifeng, China.
Tumour Biol. 2017 Jun;39(6):1010428317703825. doi: 10.1177/1010428317703825.
MicroRNA 183 (miR-183) was identified to be downregulated in nasopharyngeal carcinoma spheroids and served as a tumor suppressor in nasopharyngeal carcinoma. However, the regulatory mechanism of miR-183 and its role in cisplatin (DDP) resistance in nasopharyngeal carcinoma cells are still unclear. The expression of miR-183 and metastasis-associated protein 1 at messenger RNA and protein levels in nasopharyngeal carcinoma tissues and cells was evaluated using quantitative reverse transcription real-time polymerase chain reaction and western blotting, respectively. CNE1 and CNE2 cells were transfected with miR-183 mimic, miR-183 inhibitor, pcDNA-metastasis-associated protein 1, or respective controls. The effects of miR-183 and metastasis-associated protein 1 overexpression on cell proliferation, invasion, and DDP-induced apoptosis were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Transwell invasion assay, and flow cytometry analysis, respectively. Luciferase reporter assay was performed to explore whether miR-183 directly targeted metastasis-associated protein 1. Xenograft tumor experiment was applied to confirm the biological function of miR-183 in vivo. MiR-183 was downregulated in nasopharyngeal carcinoma tissues and cells and negatively correlated with metastasis-associated protein 1 expression. Ectopic expression of miR-183 markedly suppressed cell proliferation and invasion and strikingly enhanced DDP-induced apoptosis in nasopharyngeal carcinoma cells, whereas metastasis-associated protein 1 overexpression partially reversed these effects. Luciferase reporter assay demonstrated that metastasis-associated protein 1 was a direct target of miR-183. MiR-183 negatively regulated the expression of metastasis-associated protein 1 at both the messenger RNA and protein levels. Xenograft tumor experiment indicated that miR-183 overexpression repressed tumor growth and improved DDP-induced cytotoxicity in nasopharyngeal carcinoma cells in vivo. MiR-183 overexpression inhibited tumorigenesis and enhanced DDP-induced cytotoxicity by targeting metastasis-associated protein 1 in nasopharyngeal carcinoma, contributing to the development of novel therapeutic approaches for the treatment of clinical nasopharyngeal carcinoma patients.
微小RNA 183（miR-183）被证实在鼻咽癌球体中表达下调，并在鼻咽癌中发挥肿瘤抑制作用。然而，miR-183的调控机制及其在鼻咽癌细胞顺铂（DDP）耐药中的作用仍不清楚。分别采用定量逆转录实时聚合酶链反应和蛋白质免疫印迹法评估miR-183和转移相关蛋白1在鼻咽癌组织和细胞中信使核糖核酸和蛋白质水平的表达。用miR-183模拟物、miR-183抑制剂、pcDNA-转移相关蛋白1或各自的对照转染CNE1和CNE2细胞。分别通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐法、Transwell侵袭试验和流式细胞术分析检测miR-183和转移相关蛋白1过表达对细胞增殖、侵袭及DDP诱导凋亡的影响。进行荧光素酶报告基因试验以探究miR-183是否直接靶向转移相关蛋白1。应用异种移植瘤实验在体内证实miR-183的生物学功能。miR-183在鼻咽癌组织和细胞中表达下调，且与转移相关蛋白1的表达呈负相关。miR-183的异位表达显著抑制鼻咽癌细胞的增殖和侵袭，并显著增强DDP诱导的凋亡，而转移相关蛋白1的过表达部分逆转了这些作用。荧光素酶报告基因试验表明转移相关蛋白1是miR-183的直接靶点。miR-183在信使核糖核酸和蛋白质水平均负向调节转移相关蛋白1的表达。异种移植瘤实验表明，miR-183过表达可抑制体内鼻咽癌的肿瘤生长并增强DDP诱导的细胞毒性。miR-183过表达通过靶向转移相关蛋白1抑制鼻咽癌的肿瘤发生并增强DDP诱导的细胞毒性，有助于开发治疗临床鼻咽癌患者的新治疗方法。
