定量蛋白质组学分析SDF2L1过表达后鼻咽癌细胞中的差异蛋白

Quantitative proteomic analysis of differential proteins in nasopharyngeal carcinoma cells after SDF2L1 overexpression

目的:分析基质细胞衍生因子2样1(SDF2L1)过表达对人鼻咽癌(NPC)细胞5-8F中蛋白表达谱的影响。方法:提取SDF2L1基因过表达细胞株(5-8Fg)及空载细胞株(5-8F1)的总蛋白,利用非标记定量(LFQ)蛋白组学技术和串联质谱分析技术筛选差异表达蛋白(DEPs),应用R软件对DEPs进行GO和KEGG富集分析,Cytoscape软件筛选关键的DEPs,Oncomine数据库对关键DEPs进行验证,RT-qPCR和western blotting法对筛选出来的关键DEPs进行验证。结果:在5-8Fg细胞中共筛选出730个DEPs,其中296个DEPs上调,434个DEPs下调。富集分析结果显示,DEPs主要定位在核糖体中,参与核糖体合成和内质网蛋白加工等过程,发挥黏附蛋白和催化活性等功能。通过Cytoscape和Oncomine数据库验证出核糖体蛋白L14(RPL14)、RPS15A等9个关键DEPs。SDF2L1过表达后RPL14基因(P<0.05)表达下调。结论:SDF2L1过表达后显著改变了NPC细胞中的蛋白表达特征,RPL14可能是NPC的促癌因子。

Objective:To analyze the effect of stromal cell-derived factor 2-like 1(SDF2L1) overexpression on protein expression profiles in human nasopharyngeal carcinoma(NPC) cells 5-8F.Methods:Total proteins of SDF2L1 gene overexpression cell line(5-8Fg) and null cell line(5-8F1) were extracted,and differentially expressed proteins(DEPs) were screened using label-free qualitative(LFQ) proteomics and tandem mass spectrometry analysis.R software was used for GO and KEGG enrichment analysis of DEPs,Cytoscape software was used to screen key DEPs,Oncomine database was used to validate key DEPs,and RT-qPCR and Western blotting were used to validate the screened key DEPs.Results:A total of 730 DEPs were screened in 5-8Fg cells,of which 296 DEPs were up-regulated and 434 DEPs were down-regulated.The results of enrichment analysis showed that DEPs were mainly localized in ribosomes and involved in processes such as ribosome synthesis and endoplasmic reticulum protein processing,acting as adhesins,catalysts,etc.Nine key DEPs,including ribosomal protein L14(RPL14) and RPS15A,were verified by Cytoscape and Oncomine databases,and the expression of RPL14 gene was down-regulated after SDF2L1 overexpression(P<0.05).Conclusion:SDF2L1 overexpression significantly alters the protein expression profile in NPC cells,and RPL14 may be a pro-oncogenic factor in NPC.