过表达LncRNA IFNG-AS1急性淋巴细胞白血病细胞株的定量蛋白质组学分析  

Quantitative Proteomic Analysis of Overexpressing LncRNA IFNG-AS1 Acute Lymphoblastic Leukemia Cell Lines

作　　者：吴珊 蒋丹 徐广贤 WU Shan;JIANG Dan;XU Guangxian(Clinical School of Medicine,Ningxia Medical University,Yinchuan 750004,China;Department of Medical Inspection,School of Medical Technology,Guangdong Medical University,Dongguan 523000,China)

机构地区：[1]宁夏医科大学临床医学院,银川750004 [2]广东医科大学医学技术学院医学检验系,东莞523000

出　　处：《宁夏医科大学学报》2022年第5期479-486,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(81860355)。

摘　　要：目的通过构建IFNG-AS1过表达细胞系,寻找促进急性淋巴细胞白血病(ALL)进展的长链非编码RNA(LncRNA)IFNG-AS1的潜在靶点。方法采用IFNG-AS1过表达慢病毒和空载体组侵染ALL细胞系jurkat构建IFNG-AS1过表达的jurkat细胞,采用qRT-PCR方法检测两组细胞中IFNG-AS1的表达情况。采用高效液相色谱(HPLC)分离、串联质谱标记(TMT)定量蛋白质组学、液相色谱-质谱(LC-MS/MS)分析方法鉴定IFNG-AS1过表达jurkat细胞中差异表达蛋白(DEPs),并对其调控网络进行生物信息学分析,采用qRT-PCR对筛选的DEPs进行进一步验证。结果IFNG-AS1过表达jurkat细胞中91个蛋白表达显著上调,50个蛋白表达显著下调。DEPs与细胞分裂和免疫调节相关,具有催化活性和离子结合功能,参与了多种重要的信号通路(PPAR信号通路和PI3K/AKt信号通路)、代谢通路、DNA复制的调控,在细胞组成、生物过程和分子功能等方面具有重要作用。qRT-PCR结果显示FSIP2、PYGM、TTLL1和LIMD2的表达升高最为显著,ATP2A1、ESCO2和DNMT3B的表达降低最为显著,与蛋白质组学结果一致。结论LncRNA IFNG-AS1可影响jurkat细胞中多种蛋白的表达,且主要和TTLL1、PYGM相关。Objective By constructing IFNG-AS1 overexpressing cell lines,we searched for potential targets of long non-coding RNA(LncRNA)IFNG-AS1 that promotes the progression of acute lymphoblastic leukemia(ALL).Methods The acute lymphoblastic leukemia cell line jurkat was infected with IFNG-AS1 overexpressing lentivirus and empty vector group to construct IFNG-AS1 overexpressing jurkat cells,and qRT-PCR was used to detect the expression of IFNG-AS1 in the two groups of cells.Identification of differentially expressed proteins(DEPs)in IFNG-AS1 overexpressing jurkat cells by high performance liquid chromatography(HPLC)separation,tandem mass spectrometry tagging(TMT)quantitative proteomics,and liquid chromatography-mass spectrometry(LC-MS/MS)analysis,and bioinformatics analysis of its regulatory network was performed,and qRT-PCR was used to further verify the screened DEPs.Results 91 proteins were significantly up-regulated and 50 proteins were down-regulated in IFNG-AS1 overexpressing jurkat cells.Bioinformatics analysis showed that these DEPs were related to cell division and immune regulation,had catalytic activity and ion-binding function,and participated in a variety of important signaling pathways(PPAR signaling pathway and PI3K/AKt signaling pathway),metabolic pathways,DNA replication.It plays an important role in cell composition,biological process and molecular function.qRT-PCR verification found that the expressions of FSIP2,PYGM,TTLL1 and LIMD2 were most significantly increased,and the expressions of ATP2A1,ESCO2 and DNMT3B were most significantly decreased,which was consistent with the proteomics results.Conclusion LncRNA IFNG-AS1 can affect the expression of various proteins in jurkat cells,and it is mainly related to TTLL1 and PYGM.

关 键 词：急性淋巴细胞白血病 LncRNA IFNG-AS1 定量蛋白质组学 差异蛋白表达 

分 类 号：R392.12[医药卫生—免疫学]