猫CDH1基因在过表达c-MYC成纤维细胞中的表达变化及生物信息学分析  

Expression and Bioinformatics Analysis of Cat CDH1 Gene in c-MYC Overexpressed Fibroblasts

作　　者：安洁 杨洁 窦敏敏 孙楠楠 赵迪鹏 杜荣[2] 秦健 AN Jie;YANG Jie;DOU Minmin;SUN Nannan;ZHAO Dipeng;DU Rong;QIN Jian(College of Life Science,Shanxi Agricultural University,Taigu 030801,China;College of Veterinary Medicine,Shanxi Agricultural University,Taigu 030801,China;Center of Experiment Teaching,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学生命科学学院,太谷030801 [2]山西农业大学动物医学学院,太谷030801 [3]山西农业大学实验教学中心,太谷030801

出　　处：《中国畜牧兽医》2024年第2期443-452,共10页China Animal Husbandry & Veterinary Medicine

基　　金：山西省基础研究计划(自由探索类)自然科学研究面上项目(20210302123394);山西省回国留学人员科研资助项目(2022-102);国家自然科学基金项目(32272964、31872438);山西省留学回国人员科技活动择优资助项目;山西农业大学中青年拔尖创新人才支持计划(BJRC201204);山西农业大学横向科技项目(2015HX12)。

摘　　要：[目的]试验旨在研究骨髓细胞瘤病毒癌基因同源物(myelocytomatosis viral oncogene homolog, c-MYC)对猫成纤维细胞的影响及其与E-钙黏蛋白(cadherin 1,CDH1)基因表达和分子特性的关系,为将其应用于猫肿瘤疾病防治和组织损伤修复提供依据。[方法]通过组织贴壁法对猫胎儿成纤维细胞进行分离培养,利用电转仪将PB-TRE-c-MYC质粒转染至细胞并观察细胞形态,利用实时荧光定量PCR检测CDH1基因的表达情况,并通过生物信息学软件分析CDH1蛋白的理化性质和结构特征。[结果]过表达c-MYC导致细胞形态发生变化,从间质细胞的长梭型向上皮细胞的鹅卵石状发生转变,且使上皮细胞标志基因CDH1的表达极显著上调(P<0.01)。生物信息学分析显示,猫CDH1蛋白有881个氨基酸,其中含量最多的是亮氨酸,定位于细胞膜,存在4个CA结构域,介导细胞与细胞的接触,属于亲水性的酸性蛋白,主要由无规则卷曲组成,可能存在由Sec易位子转运并被信号肽酶Ⅰ(Sec/SPⅠ)切割的信号肽位点。[结论]猫CDH1基因的表达可被外源性重编程因子c-MYC激活,其编码的钙黏蛋白可促进猫胎儿成纤维细胞的间质-上皮转化,以抑制细胞癌化。[Objective]This study was aimed to investigate the effect of myelocytomatosis viral oncogene homolog(c-MYC)on cat fibroblasts and its relationship with cadherin 1(CDH1)gene expression and molecular characteristics,so as to provide a basis for application in the prevention and treatment of tumor diseases and the repair of tissue damage in cat.[Method]Cat fetal fibroblasts were isolated and cultured by tissue adherence,and the PB-TRE-c-MYC plasmid was transfected into the cells by electroporator,then the cell morphology was observed.The expression of CDH 1 gene was detected by Real-time quantitative PCR,and the physicochemical properties and structural characteristics of CDH1 protein were analyzed by bioinformatics software.[Result]c-MYC overexpression led to the changes in cell morphology,with a transition from the long shuttle shape of mesenchymal cells to the pebble shape of epithelial cells,and the expression of epithelial cell marker gene CDH 1 was extremely significantly upregulated(P<0.01).Bioinformatics analysis results showed that CDH1 protein in cat had 881 amino acids,of which the most abundant was leucine,localized in cell membrane,with 4 CA domains,which mediated cell-cell contact.CDH1 protein belonged to hydrophilic acidic proteins,mainly composed of random coil,and might have signaling peptide sites transported by Sec translocons and cleaved by signal peptidaseⅠ(Sec/SPⅠ).[Conclusion]The expression of CDH 1 gene in cat could be activated by exogenous reprogramming factor c-MYC,and its encoding protein cadherin could promote the mesenchymal-to-epithelial transition of cat fetal fibroblasts to inhibit cell carcinogenesis.

关 键 词：猫 C-MYC基因 CDH1基因 间质-上皮转化 

分 类 号：S829.3[农业科学—畜牧学]