Exploration of Mechanism for Meisoindigo-Inducing K562 Cell Apoptosis by Using Quantitative Proteomic Analysis.
10.7534/j.issn.1009-2137.2018.06.002
- Author:
Xin-He MAO
1
;
Ying-Xi XU
1
;
Hai-Yan XING
1
;
Zheng TIAN
1
;
Ke-Jing TANG
1
;
Lu LIU
2
;
Qing RAO
1
;
Min WANG
1
;
Jian-Xiang WANG
3
Author Information
1. State Key Laboratory of Experimental Hematology, Institute of Hematology&Blood Disease Hospital, Chinese Academy of Medical Sciences&Peking Union Medical College, Tianjin 300020, China.
2. Jingjie PTM BioLab Co Ltd, Hangzhou Economic&Technologic Development Area, Hangzhou 310018, Zhejiang Province, China.
3. State Key Laboratory of Experimental Hematology, Institute of Hematology&Blood Disease Hospital, Chinese Academy of Medical Sciences&Peking Union Medical College, Tianjin 300020, China E-mail: wangjx@ihcams.ac.cn.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Humans;
Indoles;
K562 Cells;
Proteomics;
Tandem Mass Spectrometry
- From:
Journal of Experimental Hematology
2018;26(6):1589-1597
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To screen the differentially expressed proteins at the early stage of K562 cells treated with meisoindigo by using tandem mass tags (TMT)-based proteomics technology, and to explore the mechanism for meisoindigo-inducing apoptosis.
METHODS:The half inhibitory concentration (IC) of mesoindigo on K562 cells was determined by CCK8. The flow cytometry was used to assay the apoptosis of K562 cells treated by meisoindigo or DMSO. Total proteins were extracted from the cells treated with 0.2% DMSO (control) or 20 μmol/L meisoindigo (Test) for 2 hours. Then, the TMT-labeling HPLC-MS/MS was used to identify and quantify the peptides and their abundance, all the tests were repeated for 3 times. The Mascot software was used to identify the proteins; the GO annotations, enrichment and cluster analysis were used to analyze the differentially expressed proteins.
RESULTS:Meisoindigo-induced K562 cell apoptosis in a dose-dependent manner (r=0.98), 5 544 proteins were identified, 4792 of which were quantified. The protein with expression difference>1.5-folds in Test group accoanted for 8, out of which the expression of 4 proteins were up-regulated and 4 were down-regulated. The differentially expressed proteins mainly associated with reactive oxygen species (ROS).
CONCLUSION:Several proteins including DDIT4 were found to have dramatic changes in the early stage of K562 cells treated with meisoindigo by using quantitative proteomics technology. The ROS metabolic process may play important roles in meisoindigo-inducing apoptosis of K562 cells.
