锌指蛋白ZNF750及其突变体在食管鳞癌细胞上皮间质转化中的作用及机制

项目名称： 锌指蛋白ZNF750及其突变体在食管鳞癌细胞上皮间质转化中的作用及机制

项目编号： No.81502135

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 孔鹏洲

作者单位： 山西医科大学

项目金额： 18万元

中文摘要： 食管癌是我国第五大肿瘤，鳞癌是其主要病理类型，由于缺乏有效的早期诊断和治疗方法，预后极差。肿瘤侵袭转移是食管癌患者的主要死因，上皮间质转化是其最重要机制之一。锌指蛋白750是课题组前期对食管鳞癌全基因组及外显子组测序发现的显著突变基因。实验表明ZNF750起抑癌作用，敲除和截断突变显著促进癌细胞增殖、侵袭和迁移。生物信息学提示ZNF750的抑癌作用与EMT相关，但ZNF750及其突变体在食管鳞癌EMT中的作用和机制尚不清楚。本项目拟采用生长曲线、克隆形成、侵袭实验和动物实验等检测ZNF750及其突变体在食管鳞癌EMT中的作用；采用染色质免疫沉淀、双荧光素酶报告实验、RNA干扰、挽救实验等研究ZNF750抑癌作用是否通过转录抑制EMT转录因子SNAI1/2实现，并通过荧光定位、ChIP测序等进一步研究ZNF750突变体的功能，为揭示食管癌新的癌变机理、设计新的抗肿瘤治疗方案提供理论基础。

中文关键词： 食管鳞癌；上皮间质转化；锌指蛋白750；转录抑制；snail家族锌指蛋白1/2

英文摘要： Esophageal carcinoma is the fifth-most-common cancer in China，with oesophageal squamous cell carcinoma (ESCC) being the predominant histology. And lack of effective early diagnostic and therapeutic methods leads to a poor prognosis of ESCC patients. Invasion and metastasis are the major causes of death in ESCC patients and epithelial-mesenchymal transition (EMT) plays an important role in invasion and metastasis of cancer. Our previous study of 14 whole genome sequencing and 90 exome sequencing of ESCC showed that ZNF750, a novel member of the zinc-finger transcription factors, harbors significant inactivation mutations. Bioinformatics predicted inactivation of ZNF750 may be a driver event in the development of ESCC. Further studies in vitro showed that ZNF750 may act as a tumor suppressor in ESCC and knockdown or truncating mutations of ZNF750 promote the proliferation, invasion and migration of ESCC cells. Bioinformatics indicated downregulation of ZNF750 is associated with EMT, however, the effect and molecular mechanism of ZNF750 and its mutants in EMT remain unrevealed. This proposal aims to validate the role of ZNF750 and its mutants in the process of EMT in ESCC by cell growth curve, colony formation, transwell invasion assay, zoopery, and further reveal their molecular mechanism in the transcription repression of SNAI1/2，two key EMT transcription factors, by chromatin immunoprecipitation (ChIP), dual-luciferase reporter assay, RNA interference, rescue experiment, immunofluorescence localization and ChIP-sequence. These results will facilitate early diagnosis, prevention, and treatment of ESCC in Chinese population.

英文关键词： esophageal squamous cell carcinoma;epithelial-mesenchymal transition;Zinc Finger Protein 750;transcription repression;SNAI1/2