[1]张凡,胡嘉琦,敖慧娟,等.MDCK细胞成瘤相关蛋白的筛选及验证[J].江苏农业学报,2025,(10):2009-2017.[doi:doi:10.3969/j.issn.1000-4440.2025.10.014]
　ZHANG Fan,HU Jiaqi,AO Huijuan,et al.Screening and verification of tumor-associated proteins in MDCK cells[J].,2025,(10):2009-2017.[doi:doi:10.3969/j.issn.1000-4440.2025.10.014]
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MDCK细胞成瘤相关蛋白的筛选及验证()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年10期
页码:
2009-2017
栏目:
畜牧兽医·水产养殖·益虫饲养
出版日期:
2025-10-31

文章信息/Info

Title:
Screening and verification of tumor-associated proteins in MDCK cells
作者:
张凡1; 2; 3; 4; 胡嘉琦4; 敖慧娟1; 2; 3; 4; 李睿1; 2; 3; 4; 蓝雯琳4 ; 谈笑4; 乔自林1; 2; 3; 5; 杨琨1; 2; 3; 4
(1.西北民族大学细胞基质疫苗关键技术与产业化教育部工程研究中心,甘肃兰州730030；2.西北民族大学甘肃省动物细胞技术创新中心,甘肃兰州730030；3.西北民族大学生物工程与技术国家民委重点实验室,甘肃兰州730030；4.西北民族大学生命科学与工程学院,甘肃兰州730030；5.甘肃省生物工程材料工程研究中心,甘肃兰州730030)
Author(s):
ZHANG Fan1; 2; 3; 4; HU Jiaqi4; AO Huijuan1; 2; 3; 4; LI Rui1; 2; 3; 4; LAN Wenlin4; TAN Xiao4; QIAO Zilin1; 2; 3; 5; YANG Kun1; 2; 3; 4
(1.Engineering Research Center of Key Technology and Industrialization of Cell Matrix Vaccine of Ministry of Education, Northwest Minzu University, Lanzhou 730030, China；2.Gansu Tech Innovation Center of Animal Cell, Northwest Minzu University, Lanzhou 730030, China；3.Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China；4.College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China；5.Gansu Provincial Bioengineering Materials Engineering Research Center, Lanzhou 730030, China)
关键词:
MDCK细胞; 数据非依赖性采集质谱(DIA-MS); 成瘤性; 双皮质素样激酶1(DCLK1); 组织因子途径抑制剂2(TFPI2)
Keywords:
MDCK cell; data-independent acquisition mass spectrometry (DIA-MS); tumorigenic; doublecortin-like kinase 1 (DCLK1); tissue factor pathway inhibitor 2 (TFPI2)
分类号:
Q942.6
DOI:
doi:10.3969/j.issn.1000-4440.2025.10.014
文献标志码:
A
摘要:
为明确Madin-Darby犬肾(MDCK)细胞成瘤机制,本研究基于数据非依赖性采集质谱(DIA-MS)技术,以非成瘤性MDCK细胞与高成瘤性MDCK细胞为供试材料,分析不同成瘤性MDCK细胞的蛋白质表达差异。结果表明,非成瘤性MDCK细胞与高成瘤性MDCK细胞中共鉴定到11 359个蛋白质,其中差异倍数(FC)>1.5且校正P<0.05的差异蛋白质数量为163个。非成瘤性MDCK细胞中93个差异蛋白质上调表达,70个差异蛋白质下调表达。差异蛋白质主要参与蛋白质磷酸化、胞内信号转导及肽酶活性负调控,且功能富集于细胞外空间。2个成瘤关键蛋白质双皮质素样激酶1(DCLK1)和组织因子通路抑制因子2(TFPI2)的实时荧光定量PCR和蛋白质免疫印迹(Western blot)结果显示,非成瘤性MDCK细胞中,DCLK1基因相对表达水平极显著低于成瘤性MDCK细胞,TFPI2基因相对表达水平极显著高于成瘤性MDCK细胞。本研究结果为MDCK细胞成瘤机制及肿瘤标志物筛选提供了潜在分子调控靶点。
Abstract:
In order to clarify the tumorigenic mechanism of Madin-Darby canine kidney (MDCK) cells, based on data-independent acquisition mass spectrometry (DIA-MS) technology, non-tumorigenic MDCK cells and highly tumorigenic MDCK cells were used as test materials in this study to analyze the protein expression differences of different tumorigenic MDCK cells. The results showed that a total of 11 359 proteins were identified in non-tumorigenic MDCK cells and highly tumorigenic MDCK cells, of which the number of differentially expressed proteins with fold change (FC)>1.5 and corrected P<0.05 was 163. In non-tumorigenic MDCK cells, 93 differentially expressed proteins were up-regulated and 70 differentially expressed proteins were down-regulated. Differentially expressed proteins were mainly involved in protein phosphorylation, intracellular signal transduction and negative regulation of peptidase activity, and their functions were enriched in extracellular space. Results of real-time fluorescence quantitative PCR and Western blot analysis of two key tumorigenic proteins, doublecortin-like kinase 1 (DCLK1) and tissue factor pathway inhibitor 2 (TFPI2) showed that the relative expression level of DCLK1 gene in non-tumorigenic MDCK cells was highly significantly lower than that in tumorigenic MDCK cells, and the relative expression level of TFPI2 gene was highly significantly higher than that in tumorigenic MDCK cells. The results of this study provide potential molecular regulatory targets for the tumorigenic mechanism of MDCK cells and the screening of tumor markers.

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备注/Memo

备注/Memo:
收稿日期：2025-04-11基金项目：甘肃省青年人才项目(2025QNGR63)；甘肃省高等学校教师创新基金项目(2023B-055);兰州市青年科技人才创新项目(2023-QN-70);中央高校基本科研业务费专项资金项目(31920250051、31920250071)作者简介：张凡(2002-),男,湖南张家界人,硕士研究生,主要从事生物工程研究。(Tel)15174462439;(E-mail)2495458049@qq.com通讯作者：杨琨,(E-mail)186152592@xbmu.edu.cn
更新日期/Last Update: 2025-11-17