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

doi:10.16431/j.cnki.1671-7236.2024.02.001

中国畜牧兽医 ›› 2024, Vol. 51 ›› Issue (2): 443-452.doi: 10.16431/j.cnki.1671-7236.2024.02.001

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

安洁¹, 杨洁¹, 窦敏敏², 孙楠楠¹, 赵迪鹏², 杜荣², 秦健^1,2,3

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

收稿日期:2023-08-03 出版日期:2024-02-05 发布日期:2024-01-29
作者简介:安洁,E-mail:657783994@qq.com。

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

AN Jie¹, YANG Jie¹, DOU Minmin², SUN Nannan¹, ZHAO Dipeng², DU Rong², QIN Jian^1,2,3

1. College of Life Science, Shanxi Agricultural University, Taigu 030801, China;
2. College of Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;
3. Center of Experiment Teaching, Shanxi Agricultural University, Taigu 030801, China

Received:2023-08-03 Online:2024-02-05 Published:2024-01-29
Contact: 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) E-mail:qinjian969@163.com
Supported by:
The project was supported by the National Key Research and Development Program of China (2019YFC1905301);National Natural Science Foundation of China (22078115,21776108,21690083,22008078).

摘要/Abstract

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。

关键词: 猫; c-MYC基因; CDH1基因; 间质-上皮转化

Abstract: 【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/SPⅠ).【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.

Key words: cat; c-MYC gene; CDH1 gene; mesenchymal-to-epithelial transition

中图分类号:

S829.3

安洁, 杨洁, 窦敏敏, 孙楠楠, 赵迪鹏, 杜荣, 秦健. 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[J]. 中国畜牧兽医, 2024, 51(2): 443-452.

AN Jie, YANG Jie, DOU Minmin, SUN Nannan, ZHAO Dipeng, DU Rong, QIN Jian. Expression and Bioinformatics Analysis of Cat CDH1 Gene in c-MYC Overexpressed Fibroblasts[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(2): 443-452.

参考文献

[1] GREENBURG G,HAY E D.Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells[J].Journal of Cell Biology,1982,95(1):333-339.
[2] ZAVADIL J,BITZER M,LIANG D,et al.Genetic programs of epithelial cell plasticity directed by transforming growth factor-beta[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(12):6686-6691.
[3] DONGRE A,WEINBERG R A.New insights into the mechanisms of epithelial-mesenchymal transition and implications for cancer[J].Nature Reviews Molecular Cell Biology,2019,20(2):69-84.
[4] KALLURI R,WEINBERG R A.The basics of epithelial-mesenchymal transition[J].Journal of Clinical Investigation,2009,119(6):1420-1428.
[5] BERNARDI Y,STROBL-MAZZULLA P H.What we can learn from embryos to understand the mesenchymal-to-epithelial transition in tumor progression[J].Biochemical Journal,2021,478(9):1809-1825.
[6] AKHMETKALIYEV A,ALIBRAHIM N,SHAFIEE D,et al.EMT/MET plasticity in cancer and go-or-grow decisions in quiescence:The two sides of the same coin?[J].Molecular Cancer,2023,22(1):90.
[7] ZEISBERG M,NEILSON E G.Biomarkers for epithelial-mesenchymal transitions[J].Journal of Clinical Investigation,2009,119(6):1429-1437.
[8] RADICE G L.N-cadherin-mediated adhesion and signaling from development to disease:Lessons from mice[J].Progress in Molecular Biology and Translational Science,2013,116:263-289.
[9] LUO Y,FERREIRA-CORNWELL M,BALDWIN H,et al.Rescuing the N-cadherin knockout by cardiac-specific expression of N-or E-cadherin[J].Development,2001,128(4):459-469.
[10] SHENOY S.CDH1(E-cadherin) mutation and gastric cancer:Genetics,molecular mechanisms and guidelines for management[J].Cancer Management and Research,2019,11:10477-10486.
[11] VAN ZIJL F,KRUPITZA G,MIKULITS W.Initial steps of metastasis:Cell invasion and endothelial transmigration[J].Mutation Research,2011,728(1-2):23-34.
[12] 袁少艳,何秀萍.上皮-间质转化(EMT)在上皮性卵巢癌的作用[J].中国生育健康杂志,2020,31(6):588-589. YUAN S Y,HE X P.Role of epithelial-mesenchymal transition (EMT) in epithelial ovarian cancer[J].Chinese Journal of Reproductive Health,2020,31(6):588-589.(in Chinese)
[13] GARCÍA-RUVALCABA A,VÁZQUEZ-IBARRA K C,MAGAÑA-TORRES M T,et al.Low expression of E-cadherin and CDH1 variants associated with diffuse gastric cancer[J].Revista de Investigación Clínica,2023,75(1):37-44.
[14] ZHANG Y,WEINBERG R A.Epithelial-to-mesenchymal transition in cancer:Complexity and opportunities[J].Frontiers of Medicine,2018,12(4):361-373.
[15] QIN D,LI W,ZHANG J,et al.Direct generation of ES-like cells from unmodified mouse embryonic fibroblasts by Oct4/Sox2/Myc/Klf4[J].Cell Research,2007,17(11):959-962.
[16] 石清影,朱友明.c-Myc调控lnc-CCDC117-1对舌鳞癌进展的影响[J].安徽医科大学学报,2023,58(6):1001-1010. SHI Q Y,ZHU Y M.Effect of c-Myc-regulated lnc-CCDC117-1 on the progression of tongue squamous cell carcinoma[J].Acta Universitatis Medicinalis Anhui,2023,58(6):1001-1010.(in Chinese)
[17] GAO F Y,LI X T,XU K,et al.c-MYC mediates the crosstalk between breast cancer cells and tumor microenvironment[J].Cell Communication and Signaling,2023,21(1):28.
[18] LI Y,CASEY S C,FELSHER D W.Inactivation of MYC reverses tumorigenesis[J].Journal of Internal Medicine,2014,276(1):52-60.
[19] BRUNER H C,DERKSEN P W B.Loss of E-cadherin-dependent cell-cell adhesion and the development and progression of cancer[J].Cold Spring Harbor Perspectives in Biology,2018,10(3):a029330.
[20] BERX G,VAN ROY F.Involvement of members of the cadherin superfamily in cancer[J].Cold Spring Harbor Perspectives in Biology,2009,1(6):a003129.
[21] TSAI J H,YANG J.Epithelial-mesenchymal plasticity in carcinoma metastasis[J].Genes and Development,2013,27(20):2192-2206.
[22] TAKAHASHI K,TANABE K,OHNUKI M,et al.Induction of pluripotency in human somatic cells via a transient state resembling primitive streak-like mesendoderm[J].Nature Communications,2014,5:3678.
[23] BAKIR B,CHIARELLA A M,PITARRESI J R,et al.EMT,MET,plasticity,and tumor metastasis[J].Trends in Cell Biology,2020,30(10):764-776.
[24] IVANOV D B,PHILIPPOVA M P,TKACHUK V A.Structure and functions of classical cadherins[J].Biochemistry (Moscow),2001,66(10):1174-1186.
[25] 陈振鹏,刘瑞莉,王胜,等.碱地黑牛FADS1基因克隆测序及表达分析[J].中国畜牧杂志,2023,59(5):92-98. CHEN Z P,LIU R L,WANG S,et al.Cloning sequencing and expression analysis of FADS1 gene in alkaline Black cattle[J].Chinese Journal of Animal Science,2023,59(5):92-98.(in Chinese)
[26] LIM H J,ZHUANG L,FITZGERALD R C.Current advances in understanding the molecular profile of hereditary diffuse gastric cancer and its clinical implications[J].Journal of Experimental&Clinical Cancer Research,2023,42(1):57.
[27] 刘畅晓风,方秋月,谢微嫣.RNA剪接中的SF3B1功能及其磷酸化修饰的研究进展[J].生理科学进展,2023,54(5):351-358. LIU C X F,FANG Q Y,XIE W Y.Advances in the functional role of SF3B1 in RNA splicing and its phosphorylation modifications[J].Progress in Physiological Sciences,2023,54(5):351-358.(in Chinese)

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

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