Citation: LIU Wen-Hu, YUAN Jiang-Bei, CHANG Jin-Xia. Label-free Quantitative Proteomics for Investigation of Signaling Pathways of GATA6 Regulating Trastuzumab Resistance in Gastric Cancer Cells[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(2): 187-196. doi: 10.19756/j.issn.0253-3820.191612 shu

Label-free Quantitative Proteomics for Investigation of Signaling Pathways of GATA6 Regulating Trastuzumab Resistance in Gastric Cancer Cells

1.
Research Center of Molecular Metabolomics, Xiangya Hospital Central South University, Changsha 410008, China;
2.
Department of Pharmacy, North Sichuan Medical College, Nanchong 637100, China;
3.
School of Basic Medical Sciences, North Sichuan Medical College, Nanchong 637100, China;
4.
School of Pharmaceutical Sciences and Innovative Drug Research Center, Chongqing University, Chongqing 401331, China

Received Date: 17 October 2019
Revised Date: 7 November 2019

Fund Project: This work was supported by the Applied Basic Research Program of Science and Technology of Sichuan Province, China (No. 2019YJ0378), the Project of Sichuan Province Education Department (No. 17ZB0170), the Doctoral Program of North Sichuan Medical College (No. CBY17-QD05), and the Program of Science and Technology of Nanchong, China (No. 18SXHZ0402).

Trastuzumab resistance is one of the principal causes of failure in tumor chemotherapy. Previous studies show that the DNA-binding activity of GATA6, a transcription factor, has a remarkable enhancement in trastuzumab resistant gastric cancer cells (NCI N87/R), while its correlation to resistance remains unclear. In this study, Crispr/Cas9 was employed to establish a GATA6 knock-out cell line (NCI N87R/GATA6). The signaling pathways regulated by GATA6 and related to trastuzumab resistance were investigated based on label-free quantitative proteomics combined with bioinformatics. The extracted proteins were alkylated, and digested using filter aided sample preparation (FASP), and the peptides were separated via high performance liquid chromatography and thereafter quantified by LC-MS/MS. Differentially expressed proteins were screened by fold change and student's t-test between NCI N87/R and NCI N87R/GATA6 cells. WebGestalt website was adopted for Gene ontology analysis, and GeneAnalytics was utilized for pathway enrichment analysis. The results demonstrated that GATA6 knock-out enhanced the antiproliferative effect of trastuzumab on NCI N87/R cells and suppressed their invasion ability. A total of 5792 proteins were quantified by LC-MS/MS, among which 305 proteins were up-regulated in NCI N87R/GATA6 cells while 182 ones down-regulated. Pathway enrichment analysis revealed that mitochondrial transport, apoptosis, DNA damage, glucose metabolism, pyruvate metabolism and TCA cycle and Wnt/β-catenin degradation pathways exhibited significant changes. Western blot manifested that the expression of mitochondrial dyneins OPA1 and DNM1L, apoptosis protein caspase-9, TCA metabolic enzymes SUCLG2 and MDH1, and glycogen metabolic enzyme PYGL changed significantly in NCI N87R/GATA6 cells, manifesting that GATA6 knock-out gave rise to mitochondrial dysfunction and abnormal energy metabolism, and therefore inducing the apoptosis of NCI N87R/GATA6 cells. The result implicated that inhibiting the transcriptional activity of GATA6 could be an effective strategy to reverse trastuzumab resistance in gastric cancer.
- Trastuzumab,
- Resistance,
- Label-free quantitative proteomics,
- Transcription factor GATA-6,
- Signaling pathway
1. [1]
2. [2]
3. [3]
4. [4]
5. [5]
6. [6]
7. [7]
8. [8]
9. [9]
10. [10]
11. [11]
12. [12]
13. [13]
14. [14]
15. [15]
16. [16]
17. [17]
18. [18]
19. [19]
20. [20]
21. [21]
22. [22]
23. [23]
24. [24]
25. [25]
26. [26]
27. [27]
1. [1]
  Xinyi Hong , Tailing Xue , Zhou Xu , Enrong Xie , Mingkai Wu , Qingqing Wang , Lina Wu . Non-Site-Specific Fluorescent Labeling of Proteins as a Chemical Biology Experiment. University Chemistry, 2024, 39(4): 351-360. doi: 10.3866/PKU.DXHX202310010
2. [2]
  Ruixin Liu , Feng Shi , Yanping Xia , Haibing Zhu , Jiawen Cao , Kai Peng , Chuanli Ren , Juan Li , Zhanjun Yang . Universal MOF nanozyme-induced catalytic amplification strategy for label-free electrochemical immunoassay. Chinese Chemical Letters, 2024, 35(11): 109664-. doi: 10.1016/j.cclet.2024.109664
3. [3]
  Yijian Zhao , Jvzhe Li , Yunyi Shi , Jie Hu , Meiyi Liu , Yao Shen , Xinglin Hou , Qiuyue Wang , Qi Wang , Zhiyi Yao . A label-free and ratiometric fluorescent sensor based on porphyrin-metal-organic frameworks for sensitive detection of ochratoxin A in cereal. Chinese Chemical Letters, 2025, 36(4): 110132-. doi: 10.1016/j.cclet.2024.110132
4. [4]
  Yongjie ZHANG , Bintong HUANG , Yueming ZHAI . Research progress of formation mechanism and characterization techniques of protein corona on the surface of nanoparticles. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2318-2334. doi: 10.11862/CJIC.20240247
5. [5]
  Zeyu XU , Anlei DANG , Bihua DENG , Xiaoxin ZUO , Yu LU , Ping YANG , Wenzhu YIN . Evaluation of the efficacy of graphene oxide quantum dots as an ovalbumin delivery platform and adjuvant for immune enhancement. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1065-1078. doi: 10.11862/CJIC.20240099
6. [6]
  Mi Wen , Baoshuo Jia , Yongqi Chai , Tong Wang , Jianbo Liu , Hailong Wu . Improvement of Fluorescence Quantitative Analysis Experiment: Simultaneous Determination of Rhodamine 6G and Rhodamine 123 in Food Using Chemometrics-Assisted Three-Dimensional Fluorescence Method. University Chemistry, 2025, 40(4): 390-398. doi: 10.12461/PKU.DXHX202405147
7. [7]
  Jia Zhou , Huaying Zhong . Experimental Design of Computational Materials Science Combined with Machine Learning. University Chemistry, 2025, 40(3): 171-177. doi: 10.12461/PKU.DXHX202406004
8. [8]
  Jin Yan , Chengxia Tong , Yajie Li , Yue Gu , Xuejian Qu , Shigang Wei , Wanchun Zhu , Yupeng Guo . Construction of a “Dual Support, Triple Integration” Chemical Safety Practical Education System. University Chemistry, 2024, 39(7): 69-75. doi: 10.12461/PKU.DXHX202405008
9. [9]
  Yongzhi LI , Han ZHANG , Gangding WANG , Yanwei SUI , Lei HOU , Yaoyu WANG . A two-dimensional metal-organic framework for the determination of nitrofurantoin and nitrofurazone in aqueous solution. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 245-253. doi: 10.11862/CJIC.20240307
10. [10]
  Zeyi Yan , Ruitao Liu , Xinyu Qi , Yuxiang Zhang , Lulu Sun , Xiangyuan Li , Anchao Feng . Exploration of Suspension Polymerization: Preparation and Fluorescence Stability of Perovskite Polystyrene Microbeads. University Chemistry, 2025, 40(4): 72-79. doi: 10.12461/PKU.DXHX202405110
11. [11]
  Yi Li , Zhaoxiang Cao , Peng Liu , Xia Wu , Dongju Zhang . Revealing the Coloration and Color Change Mechanisms of the Eriochrome Black T Indicator through Computational Chemistry and UV-Visible Absorption Spectroscopy. University Chemistry, 2025, 40(3): 132-139. doi: 10.12461/PKU.DXHX202405154
12. [12]
  . . University Chemistry, 2024, 39(6): 0-0.
13. [13]
  . 第41卷第6期封面和目次. Acta Physico-Chimica Sinica, 2025, 41(6): -.
14. [14]
  Jibin Miao , Changjie Mao , Baokang Jin . Exploration and Practice of Virtual and Real Combination Practical Curriculum During the Construction of the National Demonstration Center for Experimental Education: A Case Study of the National Demonstration Center for Experimental Chemistry & Chemical Engineering Education (Anhui University). University Chemistry, 2024, 39(7): 106-109. doi: 10.12461/PKU.DXHX202405021
15. [15]
  Pan Liu , Yanming Sun , Alberto J. Fernández-Carrión , Bowen Zhang , Hui Fu , Lunhua He , Xing Ming , Congling Yin , Xiaojun Kuang . Bismuth-based halide double perovskite Cs₂KBiCl₆: Disorder and luminescence. Chinese Chemical Letters, 2024, 35(5): 108641-. doi: 10.1016/j.cclet.2023.108641
16. [16]
  Zhen Liu , Zhi-Yuan Ren , Chen Yang , Xiangyi Shao , Li Chen , Xin Li . Asymmetric alkenylation reaction of benzoxazinones with diarylethylenes catalyzed by B(C₆F₅)₃/chiral phosphoric acid. Chinese Chemical Letters, 2024, 35(5): 108939-. doi: 10.1016/j.cclet.2023.108939
17. [17]
  Ruizhi Yang , Xia Li , Weiping Guo , Zixuan Chen , Hongwei Ming , Zhong-Zhen Luo , Zhigang Zou . New thermoelectric semiconductors Pb5Sb12+xBi6-xSe32 with ultralow thermal conductivity. Chinese Journal of Structural Chemistry, 2024, 43(3): 100268-100268. doi: 10.1016/j.cjsc.2024.100268
18. [18]
  Zhengzheng LIU , Pengyun ZHANG , Chengri WANG , Shengli HUANG , Guoyu YANG . Synthesis, structure, and electrochemical properties of a sandwich-type {Co₆}-cluster-added germanotungstate. Chinese Journal of Inorganic Chemistry, 2024, 40(6): 1173-1179. doi: 10.11862/CJIC.20240039
19. [19]
  Xianghe Kong , Xiaoli Liao , Zhenkun Huang , Lei Mei , Hongqing Wang , Kongqiu Hu , Weiqun Shi . Designed assembly of heterometallic cluster organic frameworks based on Th₆ cluster. Chinese Chemical Letters, 2024, 35(11): 109642-. doi: 10.1016/j.cclet.2024.109642
20. [20]
  Xin Wang , Changzhao Chen , Qishen Wang , Kai Dai . Graphene quantum dot modified Bi2MoO6 nanoflower for efficient degradation of BPA under visible light. Chinese Journal of Structural Chemistry, 2024, 43(12): 100473-100473. doi: 10.1016/j.cjsc.2024.100473

Get Citation

PDF

XML

Metrics

PDF Downloads(9)
Abstract views(834)
HTML views(88)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142