Department of Radiotherapy, Hangzhou Cancer Hospital, No. 34 Yanguanxiang, Hangzhou, China.
Department of Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jingwuweiqi Road #324, Jinan, China.
J Biochem. 2019 Nov 1;166(5):433-440. doi: 10.1093/jb/mvz057.
Nasopharyngeal carcinoma (NPC) is an important type of head and neck malignant cancer with geographical distribution. MicroRNA-449b-5p (miR-449b-5p) is related to the development of various cancers, while its function in NPC remains unknown. The present study aimed to investigate the role and target gene of miR-449b-5p in NPC. Expressions of miR-449b-5p in NPC cell lines and clinical tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was determined by MTT and colony formation assays. Migration and invasion abilities after different treatment were evaluated by wound healing and Transwell assays, respectively. Dual-luciferase reporter assay was performed to explore the relationship between miR-449b-5p and tumour protein D52 (TPD52). TPD52 expression was determined by qRT-PCR and western blot assay. miR-449b-5p was significantly downregulated in NPC cell lines and clinical tissues than the matched control. Overexpression of miR-449b-5p inhibited proliferation, migration and invasion of NPC cells. Dual-luciferase reporter assay indicated that miR-449b-5p directly targeted TPD52. Furthermore, shRNA-mediated downregulation of TPD52 rectified the promotion of cell migration and invasion by miR-449b-5p inhibition. In conclusion, the present study suggests that miR-449b-5p, as a novel tumour-suppressive miRNA against NPC, inhibits proliferation, migration and invasion of NPC cells via inhibiting TPD52 expression.
鼻咽癌(NPC)是一种具有地理分布特征的重要头颈部恶性肿瘤。微小 RNA-449b-5p(miR-449b-5p)与多种癌症的发生发展有关,但其在 NPC 中的作用尚不清楚。本研究旨在探讨 miR-449b-5p 在 NPC 中的作用及其靶基因。采用实时定量聚合酶链反应(qRT-PCR)检测 NPC 细胞系和临床组织中 miR-449b-5p 的表达。通过 MTT 和集落形成实验检测细胞增殖情况。通过划痕愈合和 Transwell 实验分别评估不同处理后细胞的迁移和侵袭能力。双荧光素酶报告实验探讨 miR-449b-5p 与肿瘤蛋白 D52(TPD52)的关系。采用 qRT-PCR 和 Western blot 检测 TPD52 的表达。与匹配的对照相比,miR-449b-5p 在 NPC 细胞系和临床组织中表达明显下调。miR-449b-5p 的过表达抑制 NPC 细胞的增殖、迁移和侵袭。双荧光素酶报告实验表明,miR-449b-5p 可直接靶向 TPD52。此外,shRNA 介导的 TPD52 下调纠正了 miR-449b-5p 抑制对细胞迁移和侵袭的促进作用。综上所述,本研究表明,miR-449b-5p 作为一种新型的 NPC 肿瘤抑制 miRNA,通过抑制 TPD52 的表达,抑制 NPC 细胞的增殖、迁移和侵袭。