B7-H3 confers stemness characteristics to gastric cancer cells by promoting glutathione metabolism through AKT/pAKT/Nrf2 pathway  

作　　者：Lu Xia Yuqi Chen Juntao Li Jiayu Wang Kanger Shen Anjing Zhao Haiyan Jin Guangbo Zhang Qinhua Xi Suhua Xia Tongguo Shi Rui Li 

机构地区：[1]Jiangsu Institute of Clinical Immunology,The First Affiliated Hospital of Soochow University,Suzhou,Jiangsu 215000,China [2]Department of Gastroenterology,The First Affiliated Hospital of Soochow University,Suzhou,Jiangsu 215000,China [3]Jiangsu Key Laboratory of Clinical Immunology,Soochow University,Suzhou,Jiangsu 215000,China [4]Jiangsu Key Laboratory of Gastrointestinal Tumor Immunology,The First Affiliated Hospital of Soochow University,Suzhou,Jiangsu 215000,China [5]Department of Oncology,The First Affiliated Hospital of Naval Military Medical University,Shanghai 200433,China [6]Department of Oncology,The First Affiliated Hospital of Soochow University,Suzhou,Jiangsu 215000,China

出　　处：《Chinese Medical Journal》2023年第16期1977-1989,共13页中华医学杂志（英文版）

基　　金：supported by Suzhou Special Project on Clinical Key Diseases Treatment Technology of Suzhou Commission of Health(No.LCZX201803);People’s Livelihood and Science and Technology project of Department of Science and Technology of Suzhou(No.SS2019059);Jiangsu Provincial Medical Key Discipline(No.ZDXK202246);Key Project of Jiangsu Provincial Health Commission(No.zd2021050);the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(No.20KJA310005);Key Project of Medical Research of Jiangsu Commission of Health(No.ZDA2020008);National Natural Science Foundation of China(No.81802843);Social Development Project of Department of Science and Technology of Jiangsu Province(No.BE2019667)

摘　　要：Background:Cancer stem-like cells(CSCs)are a small subset of cells in tumors that exhibit self-renewal and differentiation properties.CSCs play a vital role in tumor formation,progression,relapse,and therapeutic resistance.B7-H3,an immunoregulatory protein,has many protumor functions.However,little is known about the mechanism underlying the role of B7-H3 in regulating gastric cancer(GC)stemness.Our study aimed to explore the impacts of B7-H3 on GC stemness and its underlying mechanism.Methods:GC stemness influenced by B7-H3 was detected both in vitro and in vivo.The expression of stemness-related markers was examined by reverse transcription quantitative polymerase chain reaction,Western blotting,and flow cytometry.Sphere formation assay was used to detect the sphere-forming ability.The underlying regulatory mechanism of B7-H3 on the stemness of GC was investigated by mass spectrometry and subsequent validation experiments.The signaling pathway(Protein kinase B[Akt]/Nuclear factor erythroid 2-related factor 2[Nrf2]pathway)of B7-H3 on the regulation of glutathione(GSH)metabolism was examined by Western blotting assay.Multi-color immunohistochemistry(mIHC)was used to detect the expression of B7-H3,cluster of differentiation 44(CD44),and Nrf2 on human GC tissues.Student’s t-test was used to compare the difference between two groups.Pearson correlation analysis was used to analyze the relationship between two molecules.The Kaplan-Meier method was used for survival analysis.Results:B7-H3 knockdown suppressed the stemness of GC cells both in vitro and in vivo.Mass spectrometric analysis showed the downregulation of GSH metabolism in short hairpin B7-H3 GC cells,which was further confirmed by the experimental results.Meanwhile,stemness characteristics in B7-H3 overexpressing cells were suppressed after the inhibition of GSH metabolism.Furthermore,Western blotting suggested that B7-H3-induced activation of GSH metabolism occurred through the AKT/Nrf2 pathway,and inhibition of AKT signaling pathway could suppress not o

关 键 词：B7-H3 Cancer stem-like cells GLUTATHIONE Protein kinase B Nuclear factor erythroid 2-related factor 2 Gastric cancer 

分 类 号：R730[医药卫生—肿瘤]