上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2021 , Vol. 41 >Issue 4: 427 - 433

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.04.003

论著·基础研究

FBXO22调控凋亡诱导因子的分子机制及其在肿瘤细胞凋亡中的作用

  • 刘朦迪 ,
  • 潘漪莲 ,
  • 朱晓娜 ,
  • 杨烁 ,
  • 杨茜 ,
  • 余韵
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  • 1.上海交通大学医学院附属瑞金医院分子医学中心，上海 200025
    2.上海交通大学医学院附属国际和平妇幼保健院生殖遗传科，上海 200030
    3.上海交通大学医学院附属国际和平妇幼保健院产科，上海 200030
刘朦迪（1989—），女，初级技师，硕士生；电子信箱：panda8979@163.com|潘漪莲（1985—），女，住院医师，硕士；电子信箱：panyilotus@163.com。

收稿日期: 2020-07-13

  网络出版日期: 2021-05-14

基金资助

国家自然科学基金(81772936);上海交通大学医学院附属国际和平妇幼保健院院级科研基金(GFY5809);上海交通大学医学院高水平地方高校创新团队(SSMU-ZDCX20180800)

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Molecular mechanism of apoptosis-inducing factor regulated by FBXO22 and its role in cancer cell apoptosis

  • Meng-di LIU ,
  • Yi-lian PAN ,
  • Xiao-na ZHU ,
  • Shuo YANG ,
  • Qian YANG ,
  • Yun YU
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  • 1.Molecular Medicine Center, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    2.Department of Reproductive Genetics, International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China
    3.Department of Obstetrics, International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China

Received date: 2020-07-13

  Online published: 2021-05-14

Supported by

National Natural Science Foundation of China(81772936);Foundation of International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine(GFY5809);Innovative Research Team of High-Level Local Universities in Shanghai(SSMU-ZDCX20180800)

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摘要

目的·挖掘F-box蛋白22（F-box only protein 22，FBXO22）的底物并明确其作用机制，进一步探讨FBXO22在肿瘤细胞凋亡中的作用。方法·利用免疫沉淀联合质谱分析寻找并验证FBXO22的相互作用蛋白凋亡诱导因子（apoptosis-inducing factor，AIF）。采用变性条件下泛素化修饰实验检测FBXO22对AIF泛素化修饰的影响。通过短发夹RNA敲低FBXO22、过表达FBXO22检测AIF的表达水平。利用流式细胞仪检测FBXO22敲低或过表达对结肠癌细胞凋亡的影响。结果·免疫沉淀联合质谱分析显示FBXO22可与AIF结合，变性条件下泛素化修饰实验显示FBXO22可介导AIF的泛素化修饰。敲低FBXO22可上调AIF水平，过表达FBXO22可下调AIF水平。流式细胞术结果显示，敲低FBXO22可促进由甲基硝基亚硝基胍（N-methyl-N'-nitro-N-nitrosoguanidine，MNNG）等诱导的结肠癌细胞凋亡，过表达FBXO22则可抑制MNNG等诱导的结肠癌细胞凋亡。结论·FBXO22通过泛素化修饰降解AIF进而调控由AIF介导的结肠癌细胞凋亡。

关键词： F-box蛋白22; 凋亡诱导因子; 泛素化修饰; 细胞凋亡

本文引用格式

刘朦迪 , 潘漪莲 , 朱晓娜 , 杨烁 , 杨茜 , 余韵 . FBXO22调控凋亡诱导因子的分子机制及其在肿瘤细胞凋亡中的作用[J]. 上海交通大学学报（医学版）, 2021 , 41(4) : 427 -433 . DOI: 10.3969/j.issn.1674-8115.2021.04.003

Abstract

Objective

·To explore the substrate of F-box only protein 22 (FBXO22) and its mechanism, and further study the role of FBXO22 in cancer cell apoptosis.

Methods

·The interaction protein of FBXO22, namely apoptosis-inducing factor (AIF), was searched and identified by immunoprecipitation (IP)-coupled liquid chromatograph mass spectrometer/mass spectrometer (LC-MS/MS) technology. The effect of FBXO22 on ubiquitination of AIF was detected by ubiquitin modification assay under denatured conditions. Short hairpin RNA (shRNA) knockdown FBXO22 and overexpression FBXO22 were used to detect the expression of AIF. The effect of FBXO22 knockdown or overexpression on apoptosis of colon cancer cells was detected by flow cytometry.

Results

·IP-coupled LC-MS/MS technology showed that FBXO22 could interact with AIF, and ubiquitin modification assay under denaturing conditions showed that FBXO22 could mediate ubiquitination of AIF. Knockdown of FBXO22 could up-regulate the level of AIF, while overexpression of FBXO22 could down-regulate the level of AIF. The results of flow cytometry showed that knockdown of FBXO22 could promote the apoptosis of colon cancer cells induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), while overexpression of FBXO22 could inhibit the apoptosis of colon cancer cells induced by MNNG.

Conclusion

· FBXO22 regulates AIF-mediated apoptosis of colon cancer cells by ubiquitination and degradation of AIF.

Key words： F-box only protein 22 (FBOX22); apoptosis-inducing factor (AIF); ubiquitination modification; cell apoptosis

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