Mechanism Of ANXA2 Regulates Triple Negative Breast Cancer Invasion And Metastasis Via Hippo Signaling Pathway

Objective:To observe the expression of annexin A2(ANXA2)in Breast cancer(BC)and its relationship with prognosis of patients,and to confirm that overexpression/ANXA2 knockdown can affect the invasion and metastasis of Triple negative Breast cancer(TNBC)cells.Furthermore,the molecular mechanism of Hippo/YAP signaling axis participating in the regulation of TNBC invasion and metastasis was further discussed.Methods:(1)The expression of ANXA2 in BC and its different molecular types in cancer Genome Atlas(TCGA)and genotypic tissue expression(GTEx)databases were analyzed using GEPIA2 online tool.(2)Kaplan-Meier Plotter database was used to analyze the association between ANXA2 and overall survival(OS)and relapse-free survival(RFS).(3)Human ANXA2 gene overexpression and knockdown virus was transfected with TNBC cell line MDA-MB-231.The changes of cell invasion and metastasis were observed by Transwell assay,and the changes of cell migration were observed by scratch assay.(4)MDA-MB-231 cells were treated with MST1/2 inhibitor XMU-MP-1,and the recovery of cell migration and invasion ability was observed by Transwell assay and scratch assay.(5)Western blot and qPCR were used to verify the changes of YES-associated protein(YAP)and gene expression after overexpression/knockout of ANXA2.Results:(1)Bioassay results showed that ANXA2 was highly expressed in breast cancer,triple negative breast cancer,HER2-positive breast cancer,and LuminalA type breast cancer,and the overall survival(OS)and relapse-free survival(RFS)of TNBC patients with high ANXA2 expression were significantly decreased(P<0.05).(2)Transwell results showed that the migration and invasion ability of MDA-MB-231 cells was significantly decreased after ANXA2 knockout(P<0.05),while the migration and invasion ability of MDA-MB-231 cells was enhanced after ANXA2 overexpression(P<0.05).Scratch test results showed that ANXA2 knockdown decreased cell migration ability(P<0.05),and ANXA2 overexpression increased cell migration ability(P<0.05).(3)Treatment of ANXA2 knockout cells with MST1/2 inhibitor XMU-MP-1 significantly restored the ability of migration and invasion(P<0.05).(4)qPCR results showed that ANXA2 knockout decreased the expression of YAP gene(P<0.05),but ANXA2 overexpression increased the expression of YAP gene(P<0.05).Western blot results showed that ANXA2 knockdown decreased the expression of YAP gene(P<0.05).Conclusion:(1)In TNBC,overall survival and relapse-free survival were reduced in patients with high ANXA2 expression.(2)The invasion and migration abilities of TNBC cells were significantly increased/decreased after overexpression/ANXA2 knockout.(3)In TNBC cells,ANXA2 regulates the invasion and metastasis of TNBC cells via the YAP-mediated Hippo signaling pathway and promotes tumor progression.