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
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)
