HSP90功能性抗体对胃癌干细胞干性特征作用的研究OA北大核心

Objective:To investigate the role of the functional antibody heat shock protein 90(HSP90)on stemness character-istics of gastric cancer stem cells and to preliminarily explore the related mechanisms.Methods:Using serum-free suspension cul-ture of gastric cancer HGC27 cells to obtain cytosphere cells(Sphere cells);treating Sphere cells with HSP90 functional antibody 9B8 and using CCK-8 assay to detect the role of 9B8 on gastric cancer stem cell resistance and proliferation;using Trans-well chamber to detect the role of 9B8 on migration and invasion of gastric cancer stem cells;using methylcellulose sphere-forming as-say to detect the role of 9B8 on the self-renewal of gastric cancer stem cells;using flow cytometry to detect the role of 9B8 on the expression of ALDH and CD44 in gastric cancer stem cells;using immunofluorescence to detect the expression of HSP90 in gas-tric cancer stem cells,and Western blot to detect the expression of HSP90,stemness-associated proteins(SOX2 and OCT4),drug-resistance-associated proteins(TOP2A and ABCG2)in gastric cancer stem cells,and epithelial-mesenchymal transition(EMT)-related proteins(Vimentin,Snail,and E-cadherin),as well as the expression of proteins related to PI3K/AKT/mTOR signaling pathway.Results:The expression of HSP90,ALDH,and CD44 in gastric cancer stem cells was higher than that of pa-rental cells,and their proliferation,drug resistance,migration,invasion,and self-renewal abilities were significantly stronger than those of parental cells(P<0.01).There was no difference in the proliferation of gastric cancer stem cells in the HSP90 functional antibody between the 9B8-treated group and the control group(P>0.05),but the migration,invasion,drug resistance,and self-renewal abilities were significantly lower than those in the control group(P<0.01),and the proportion of ALDH and CD44 ex-pression was significantly decreased(P<0.01).The expressions of HSP90,ABCG2,TOP2A,Vimentin,Snail,PI3K,p-PI3K,AKT,p-AKT,mTOR,and p-mTOR were decreased in the HSP90 functional antibody-treated group,and the expression of E-cadherin was increased.Conclusion:HSP90 functional antibody 9B8 may affect the expression of gastric cancer stem cell-re-lated stemness genes/proteins,drug resistance genes/proteins,and EMT-related genes/proteins by inhibiting the activity of AKT/PI3K/mTOR pathway,which in turn affects the stemness characteristics of gastric cancer stem cells,such as self-renewal,drug resistance,invasion,and migration,and thus provides a new way of thinking for the targeting therapy of gastric cancer stem cells.