色谱

色谱 ›› 2020, Vol. 38 ›› Issue (2): 162-168.DOI: 10.3724/SP.J.1123.2019.08013

• 特别策划：第22届全国色谱学术报告会优秀论文专栏 • 上一篇    下一篇

基于液相色谱-质谱联用的代谢组学技术研究Pokemon诱导的胞内脂代谢变化

金一宝1, 李上富2, 王珏1, 高丹2, 刘红霞2, 蒋宇扬2, 殷果1, 王铁杰1,*()   

  1. 1 深圳市药品检验研究院, 深圳市药品质量标准研究重点实验室, 广东 深圳 518057
    2 清华大学深圳研究生院, 广东 深圳 518055
  • 收稿日期:2019-08-14 出版日期:2020-02-08 发布日期:2020-12-10
  • 通讯作者: 王铁杰
  • 作者简介:王铁杰.Tel:(0755)26031728, E-mail:szyjwtj@163.com
  • 基金资助:
    中国博士后基金(2015M572370);深圳市基础研究项目(JCYJ2016053017352098)

A liquid chromatography-mass spectrometry-based metabolomic study on the changes of intracellular lipid metabolism regulated by Pokemon

JIN Yibao1, LI Shangfu2, WANG Jue1, GAO Dan2, LIU Hongxia2, JIANG Yuyang2, YIN Guo1, WANG Tiejie1,*()   

  1. 1 Shenzhen Institute for Drug Control, Shenzhen Key Laboratory of Drug Quality Standard Research, Shenzhen 518057, China
    2 Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China
  • Received:2019-08-14 Online:2020-02-08 Published:2020-12-10
  • Contact: WANG Tiejie
  • Supported by:
    Chinese Postdoctoral Science Foundation(2015M572370);Shenzhen Fundamental Research Program(JCYJ2016053017352098)

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摘要：

Pokemon是一种转录抑制因子，能够通过影响染色质的重组或直接与抑癌基因结合而抑制抑癌基因的转录，促使肿瘤形成。该文利用基于液相色谱-质谱联用的代谢组学技术研究了Pokemon在肝癌中调控细胞代谢的作用机制。通过脂质转染，获得了Pokemon高表达的HL7702细胞，分别收集转染后不同时间点的细胞。利用基于液相色谱-质谱联用技术的代谢组学方法，分析胞内代谢物的成分。根据多元统计分析的结果选出差异显著的候选代谢物，通过数据库（METLIN和HMDB）检索、二级图谱比对进行结构解析，确证了36种代谢物。通过KEGG数据库检索发现这些代谢物主要与脂质合成相关。进一步分析发现脂质合成途径中乙酰辅酶羧化酶和脂肪酸合成酶均被激活。结果显示，Pokemon可通过激活细胞中脂质合成通路而影响细胞的代谢。

关键词: 液相色谱-质谱联用, Pokemon, 代谢组学, 脂质合成

Abstract:

Pokemon, a transcriptional repressor, inhibits the transcription of tumor suppressor genes and promotes tumor formation by affecting chromatin recombination or binding directly to the tumor suppressor genes. In this study, the mechanism behind the Pokemon regulation of liver cancer cell metabolism was evaluated using liquid chromatography-mass spectrometry (LC-MS)-based metabolomics. Pokemon-overexpressing HL7702 cells were obtained by lipofection. The cells were collected at different time points after transfection and their intracellular metabolites were analyzed using an LC-MS-based metabolomic method. Based on the results of a multivariate statistical analysis, candidate metabolites with significant differences were selected, and 36 of them were confirmed by database and MS/MS analysis. These metabolites were primarily involved in lipid synthesis by KEGG database searches. Further analysis indicated that both the acetyl-coenzyme carboxylase and the fatty acid synthase were activated in the lipid synthesis pathway. The results show that Pokemon can affect cell metabolism by activating the lipid synthesis pathways in cells.

Key words: liquid chromatography-mass spectrometry (LC-MS), Pokemon, metabolomics, lipid synthesis