首页|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

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

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[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(cadherin1，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/SP Ⅰ)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。
Expression and Bioinformatics Analysis of Cat CD H1 Gene in c-MYC Overexpressed Fibroblasts
[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 CDH1 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 CDH1 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 peptidase Ⅰ(Sec/SP Ⅰ).[Conclusion]The expression of CDH1 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.

catc-MYC geneCDH1 genemesenchymal-to-epithelial transition

安洁、杨洁、窦敏敏、孙楠楠、赵迪鹏、杜荣、秦健

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山西农业大学生命科学学院,太谷 030801

山西农业大学动物医学学院,太谷 030801

山西农业大学实验教学中心,太谷 030801

猫 c-MYC基因 CDH1基因 间质-上皮转化

山西省基础研究计划(自由探索类)自然科学研究面上项目山西省回国留学人员科研资助项目国家自然科学基金国家自然科学基金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山西农业大学横向科技项目

202103021233942022-1023227296431872438BJRC2012042015HX12

2024

10.16431/j.cnki.1671-7236.2024.02.001

中国畜牧兽医
中国农业科学院北京畜牧兽医研究所

中国畜牧兽医

CSTPCD北大核心
影响因子：0.72
ISSN：1671-7236
年,卷(期)：2024.51(2)
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