Cheng Chao, Li Wenmin, Peng Xuemei, Liu Xiong, Zhang Ziyan, Liu Zhen, Deng Tongyuan, Luo Rongcheng, Fang Weiyi, Deng Xiaojie
Department of Pediatric Otolaryngology, Shenzhen Hospital, Southern Medical University, Shenzhen, Guangdong, China.
Cancer Center, Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.
Mol Ther Methods Clin Dev. 2021 Feb 4;20:615-624. doi: 10.1016/j.omtm.2021.02.001. eCollection 2021 Mar 12.
Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in Chinese and other Southeast Asians. We aimed to explore the precise mechanism for NESG1 in NPC for understanding the pathogenesis of NPC. Transwell, Boyden assays, and wounding healing were respectively performed for cell metastasis. The microRNA (miRNA) microarray and luciferase reporter assays were designed to clarify NESG1-modulated miRNAs and miR-1254-targeted protein. Western blotting assays examined the pathways regulated by miR-1254, the (Hepatoma-Derived Growth Factor) HDGF/DDX5 complex, and NESG1. The chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), and co-immunoprecipitation (coIP) assays were used to explore the DNA-protein complex and protein-protein complex. NESG1 suppressed NPC migration and invasion via Wnt/β-catenin signaling. Further, miR-1254 was confirmed as a positive downstream modulator of NESG1 reducing metastatic abilities of NPC cells and . Transduction of HDGF significantly restored cell migration and invasion ability in miR-1254-overexpressing NPC cells. In clinical samples, miR-1254 expression was negatively correlated with HDGF and positively correlated with NESG1 expression. miR-1254 acts as an independent prognostic factor for NPC, which was induced by NESG1 to suppress NPC metastasis via inactivating Wnt/β-catenin pathway and its downstream EMT signals.
鼻咽癌（NPC）是中国及其他东南亚地区最常见的恶性肿瘤之一。我们旨在探究NESG1在鼻咽癌中的精确机制，以了解鼻咽癌的发病机制。分别采用Transwell实验、Boyden实验和伤口愈合实验检测细胞转移情况。设计微小RNA（miRNA）微阵列和荧光素酶报告基因实验以阐明NESG1调控的miRNA以及miR-1254靶向的蛋白。蛋白质免疫印迹实验检测miR-1254、（肝癌衍生生长因子）HDGF/DDX5复合物和NESG1调控的信号通路。采用染色质免疫沉淀（ChIP）实验、电泳迁移率变动分析（EMSA）实验和免疫共沉淀（coIP）实验探究DNA-蛋白质复合物和蛋白质-蛋白质复合物。NESG1通过Wnt/β-连环蛋白信号通路抑制鼻咽癌的迁移和侵袭。此外，miR-1254被证实为NESG1的正向下游调节因子，可降低鼻咽癌细胞的转移能力。HDGF转导显著恢复了miR-1254过表达的鼻咽癌细胞的迁移和侵袭能力。在临床样本中，miR-1254表达与HDGF呈负相关，与NESG1表达呈正相关。miR-1254作为鼻咽癌的独立预后因素，由NESG1诱导，通过使Wnt/β-连环蛋白通路及其下游上皮-间质转化信号失活来抑制鼻咽癌转移。
