南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (10): 1191-.doi: 10.12122/j.issn.1673-4254.2019.10.10

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SETD2基因敲除前后鼻咽癌细胞中定量蛋白组学研究及差异信号富集

曾玉梅，王思思，封慕茵，邵钟铭，袁建玲，申志华，揭伟   

  • 出版日期:2019-10-20 发布日期:2019-10-20

Quantitative proteomics and differential signal enrichment in nasopharyngeal carcinoma cells with or without SETD2 gene knockout

  • Online:2019-10-20 Published:2019-10-20

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摘要： 目的分析甲基转移酶SETD2 表达改变对人鼻咽癌（NPC）细胞中蛋白表达谱的影响并富集差异信号通路。方法提取 SETD2基因敲除细胞株CNE1SETD2-KO和野生型细胞CNE1WT的总蛋白，利用Tandem Mass Tag（TMT）标记蛋白质定量技术和串联 质谱分析技术筛选出差异表达蛋白质，应用GO数据库对差异表达蛋白质进行注释和富集，应用KEGG数据库进行差异蛋白质 涉及信号通路富集。结果以表达倍数（FC）≥1.2倍且P<0.05为筛选标准，SETD2基因敲除的CNE1细胞鉴别出2049个差异表 达蛋白质，其中904个表达上调，1145个表达下调。GO功能注释结果表明，SETD2敲除后在生物过程（细胞过程及调节，细胞运 动、代谢过程及细胞组分的生物合成）、分子功能（催化活性及分子结合、转录因子活性）和细胞组分（胞膜、细胞器、大分子复合 物）等具有特征性。KEGG信号富集结果表明SETD2 敲除主要影响与肿瘤关系密切的信号有MAPK、PI3K-Akt、Ras、Rap1、 mTOR、Hippo、HIF-1、Wnt、AMPK、FoxO、ErbB、p53、JAK-STAT等。结论SETD2敲除后显著改变了NPC细胞中蛋白质表达特 征并影响多条与肿瘤关系密切的信号通路，研究结果为NPC的发病机制和治疗标靶筛选提供了参考。

Abstract: Objective To analyze the effects of alterations in the expressions of methyltransferase SETD2 on protein expression profiles in human nasopharyngeal carcinoma (NPC) cells and enrich the differential signaling pathways. Methods The total protein was extracted from SETD2-knockout cell line CNE1SETD2-KO and the wild-type cell line CNE1WT, and the differentially expressed proteins were screened by tandem mass tag (TMT) labeled protein quantification technique and tandem mass spectrometry. GO analysis was used to annotate and enrich the differentially expressed proteins, and the KEGG database was used to enrich and analyze the pathways of the differential proteins. Results With a fold change (FC)≥1.2 and P<0.05 as the screening standard, 2049 differentially expressed proteins were identified in CNE1SETD2-KO cells, among which 904 were up-regulated and 1145 were down-regulated. GO functional annotation results indicated that SETD2 knockout caused characteristic changes in multiple biological processes (cell processes and regulation, cell movement, metabolic processes, and biosynthesis of cellular components), molecular functions (catalytic activity and molecular binding, transcription factor activity), and cellular components (cell membrane, organelle, macromolecular complex). KEGG analysis showed that the differentially expressed proteins were involved in an array of signaling pathways closely related to tumors, including MAPK, PI3K-Akt, Ras, Rap1, mTOR, Hippo, HIF-1, Wnt, AMPK, FoxO, ErbB, P53 and JAK-STAT. Conclusion SETD2 knockout significantly changes the protein expression characteristics of NPC cells and affects a number of signal pathways closely related to tumors. The results provide evidence for investigation of the pathogenesis and therapeutic target screening of NPC.