Citation: CHANG Jin-Xia, WANG Yi, ZHANG Fan, LIU Wen-Hu. Proteomic Study of Transcription Factors in Trastuzumab-Resistant Gastric Cancer Based on Liquid Chromatography-Mass Spectrometry Technique[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(7): 1035-1044. doi: 10.19756/j.issn.0253-3820.191093 shu

Proteomic Study of Transcription Factors in Trastuzumab-Resistant Gastric Cancer Based on Liquid Chromatography-Mass Spectrometry Technique

CHANG Jin-Xia ^1, ,
WANG Yi ² ,
ZHANG Fan ³ ,
LIU Wen-Hu ^2,3

1.
School of Basic Medical Sciences, North Sichuan Medical College, Nanchong 637100, China;
2.
State Key Laboratory of Proteomics, National Center for Protein Sciences, Beijing 102206, China;
3.
Department of Pharmacology, North Sichuan Medical College, Nanchong 637100, China

Received Date: 13 February 2019
Revised Date: 10 May 2019

Fund Project: This work was supported by the Applied Basic Research Programs of Science and Technology of Sichuan, China (No. 2019YJ0378), the Doctoral Program of Sichuan Medical College (No. CBY17-QD05) and the Program of Science and Technology of Nanchong, China (No. 18SXHZ0402).

A method was developed for investigation of regulatory role of transcription factors (TFs) in trastuzumab resistance of gastric cancer cells by liquid chromatography-mass spectrometry coupled with concatenated tandem array of transcription factor response elements (catTFRE) pull down proteomic technique. A synthetic DNA containing catTFRE was used as an affinity reagent, on which a biotin was labeled to become a DNA decoy, and then TFs were enriched via catTFRE pull down and detected by liquid chromatography-mass spectrometry. Intensity based absolute quantification (iBAQ) was applied to quantify TFs, and the TFs with a significant change in DNA binding activity were filtered out. Signaling pathways regulated by TFs, family classification, regulatory network and TFs-targets analysis were done based on WebGestalt (2017) database. The result showed that, 359 TFs were detected and quantified, and comparing with parental cells, 61 TFs in the trastuzumab resistance gastric cancer cells showed a remarkable activity change in DNA binding, of which 48 TFs showed increased activity and 13 showed decreased activity. The activated TFs belonged to bZIP, bHLH, homebox, HMG box, Zine finger, etc. KEGG pathway enrichment analysis demonstrated that multiple pathways including cancer, MAPK, Wnt, TGF-β, apoptosis changed prominently in trastuzumab resistant gastric cells, TFs-targets analysis revealed that TFs including TCF7L2, TCF7, FOXC1, JUN, MYC, FOS, ELK4, NFκB2, DDIT3 and ATF2 had an important regulatory effect on epithelial-mesenchymal transformation (EMT), Wnt and MAPK signaling pathways which contributed to trastuzumab-resistance in gastric cancer cells. This implicated that the targeting signaling pathways regulated by these TFs could possibly reverse trastuzumab resistance in gastric cancer.
- Trastuzumab,
- Resistance,
- Concatenated tandem array of transcription factor response elements,
- Pull down,
- Proteomics
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