SHP-2 overexpression in tumor-associated macrophage accelerates angiogenesis by activating the PI3K/mTOR pathway and promotes breast cancer progression
HE Guang
JI Dong
SONG Cuiping
DOU Yalin
CAO Heju
Abstract：Objective To investigate whether Src homology 2 domain-containing protein tyrosine phosphatase(SHP-2)overexpression in tumor-associated macrophages could accelerate angiogenesis by activating the Phosphatidylinositol-3-kinase(PI3K)/mammalian target of Rapamycin(mTOR)pathway,so as to promote breast cancer progression.Methods LipofectamineTM2000 was used to establish SHP-2 overexpressing cells in the breast cancer cell line MCF-7.Twenty BALB/C nude mice were randomly divided into two groups:①SHP-2-NC group,②SHP-2+PHPS1 inhibitor group.PHPS1 inhibitors were added to the SHP-2+PHPS1 inhibitor group.The above grouped cell solution was injected into the nude mouse in the right axilla subcutaneously to construct a tumor model,and the tumor size was measured and recorded daily.Transfected THP-1 cells were treated with a 740Y-P agonist,and divided into two groups:③SHP-2-NC+740Y-P agonist group,④SHP-2+PHPS1 inhibitor+740Y-P agonist group.740Y-P agonists were added to the SHP-2-NC+740Y-P agonist group and SHP-2+PHPS1 inhibitor+740Y-P agonist group.Western blot was used to measure the expression levels of p-SHP-2,p-PI3K,p-mTOR,VEGF,COX-2,MMP2 and MMP9 protein in MCF-7 cells,respectively.Tube formation assay was used to detect tube-forming capacity of HUVEC cells.Cell scratch assay and Transwell assay were used to detect migration invasion of MCF-7 cells.Results The results of animal experiments showed that the tumor volume in the SHP-2+PHPS1 inhibitor group decreased significantly(P＜0.05)compared with the SHP-2-NC group.The results of MCF-7 cell Western blot showed that compared with the SHP-2-NC group,the levels of p-SHP-2,p-PI3K,p-mTOR,VEGF,COX-2,MMP2 and MMP9 proteins in the SHP-2+PHPS1 inhibitor group were significantly decreased(P＜0.05).Compared with the SHP-2-NC+740Y-P agonist group,the levels of p-SHP-2 proteins in the SHP-2+PHPS1 inhibitor+740Y-P agonist group were significantly decreased(P＜0.05),and there was no significant difference in p-PI3K,p-mTOR,VEGF,COX-2,MMP2 and MMP9 protein expression(P＞0.05).Compared with group①②,the level of p-PI3K,p-mTOR,VEGF,COX-2,MMP2 and MMP9 protein in group ③④ was significantly increased(P＜0.05).The results of HUVEC cells tube formation assay showed compared with the SHP-2-NC group,the number of lumen formation in the SHP-2+PHPS1 inhibitor group were significantly decreased(P＜0.05),and there was no significant difference in lumen formation number between the SHP-2-NC+740Y-P agonist group and the SHP-2+PHPS1 inhibitor+740Y-P agonist group(P＞0.05).Compared with the ①② group,the number of lumen formation increased significantly in the ③④ group(P＜0.05).Among the four groups of MCF-7 cells,the migration invasion capacity of the SHP-2-NC group was greater than those in the SHP-2+PHPS1 inhibitor group(P＜0.05),and there was no significant difference between SHP-2+PHPS1 inhibitor and SHP-2+PHPS1 inhibitor+740Y-P agonist(P＞0.05).The migration invasion capacity of the ③④group was greater than those in the ①② group(P＜0.05).Conclusions Overexpression of SHP-2 in tumor-associated macrophages can accelerate angiogenesis by activating the PI3K/mTIR pathway and thereby promote breast cancer progression.
Keywords：Tumor-associated macrophagesSHP-2PI3K/mTORAngiogenesisBreast cancer
Publication Date：2025-06-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 291-298 )