基于串联质量标签定量质谱法的胶质瘤外泌体蛋白组差异分析

裴顺琪; 谭舟

doi:10.16495/j.1006-3757.2023.01.010

基于串联质量标签定量质谱法的胶质瘤外泌体蛋白组差异分析

doi: 10.16495/j.1006-3757.2023.01.010

裴顺琪^{1, 2,},
谭舟^1, ,

1.
杭州师范大学生命与环境科学学院，浙江杭州　311121
2.
浙江大学生命科学学院，浙江杭州　310058

基金项目: 高层次留学回国人员在杭创新创业项目

详细信息

作者简介:
裴顺琪（1994−），男，博士生，主要从事胶质发育及脑肿瘤研究，E-mail：shunqi_pei@qq.com

通讯作者:
谭舟（1982−），男，副教授，主要从事神经发育与疾病研究，E-mail：tanzhou@hznu.edu.cn

中图分类号: O657. 63
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- 被引次数: 0
出版历程
- 收稿日期: 2023-01-31
- 录用日期: 2023-03-06
- 修回日期: 2023-03-06
- 刊出日期: 2023-03-31

Exploration of Exosome Component Differences in Gliomas Based on Tandem Mass Tag Mass Spectrometry

PEI Shunqi^{1, 2
,},
TAN Zhou^{1
, ,}

1.
College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 311121, China
2.
College of Life Sciences, Zhejiang University, Hangzhou 310058, China

Funds: High-Level Students Returning to China (Team) Project in Hangzhou 2017

摘要

摘要: 为筛选胶质瘤外泌体中参与肿瘤迁移的差异蛋白，寻找可能抑制肿瘤细胞迁移的作用靶点. 本试验分别对具有不同迁移能力的A172、U251胶质瘤细胞系培养上清液进行了差异性探究. 结果显示，A172条件培养基具有更强的迁移促进作用，而外泌体为促进肿瘤迁移的主要成分. 随后利用串联质量标签（tandem mass tag，TMT）定量质谱法对其外泌体进行的蛋白质组学分析发现，差异蛋白（倍数改变≥2）主要富集于核糖体、间隙连接、泛素介导的蛋白降解三个KEGG（kyoto encyclopedia of genes and genomes）信号通路. 通过分析共获得了50种具有显著表达差异的外泌体蛋白，可作为胶质瘤的外泌体检测标志物及抑制肿瘤细胞迁移的潜在作用靶点.
- 胶质瘤 /
- 迁移 /
- 外泌体 /
- 串联质谱标签定量质谱法
Abstract: In order to screen for differential proteins involved in tumor migration in glioma exosomes and to find targets that may inhibit tumor cell migration, the culture supernatant discrepancies of A172 and U251 glioma cell lines with different migration capabilities was explored. The results showed that A172-conditioned medium had a stronger migration-promoting effect, while exosomes were the main components to promote tumor migration. Subsequently, the proteomic analysis of exosomes using tandem mass tag (TMT) quantitative mass spectrometry showed that differential proteins (fold change≥2) were mainly enriched in three KEGG (kyoto encyclopedia of genes and genomes) signaling pathways: ribosomes, gap junctions, and ubiquitin-mediated protein degradation. A total of 50 exosome proteins with significant expression differences were obtained, which could be used as exosome detection markers for gliomas and potential targets for inhibiting tumor cell migration.
- gliomas /
- migration /
- exosome /
- tandem mass tag mass spectrometry

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图 1 两种胶质瘤细胞（A172，U251）与NHA形态及生长差异图

（A）A172于20倍物镜下明场图，比例尺=200 μm，（B）U251于20倍物镜下明场图，比例尺=200 μm，（C）两种细胞系（A172，U251）与NHA的生长图，纵坐标代表相对吸光度

Figure 1. Morphology and growth difference between two types of glioblastoma cells (A172, U251) and NHA

下载: 全尺寸图片幻灯片

图 2 不同肿瘤细胞条件培养基对A172细胞迁移速率的影响

（A）0、24、48、72 h时的细胞迁移明场图，比例尺=500 μm，（B）图（A）细胞迁移距离的统计结果

Figure 2. Effect of different conditional mediums of tumor cells on migration rate of A172 cells

下载: 全尺寸图片幻灯片

图 3 肿瘤细胞外泌体及上清液对A172细胞迁移速率的影响

（A）0、12、24、36 h时的细胞迁移明场图，比例尺=500 μm，（B）图（A）细胞迁移距离的统计结果

Figure 3. Effect of different extracellular vesicles and supernatant on migration rate of A172 cells

下载: 全尺寸图片幻灯片

图 4 外泌体蛋白基因集富集分析图

（A）核糖体（KEGG_ribosome），（B）间隙连接（KEGG_gap junction），（C）泛素介导的蛋白降解（KEGG_ubiquitin mediated proteolysis）

Figure 4. Gene set enrichment analysis of exosomal proteins

下载: 全尺寸图片幻灯片

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