Effect Of TEAD3 On Glioma Proliferation,Migration,Invasion And Angiogenesis

BackgroundGlioma is the most common primary tumor of the central nervous system,of which high-grade glioma is the most common,accounting for more than 70%of gliomas.Due to the characteristics of high invasiveness and high migration,the tumor often remains after conventional surgical treatment supplemented with chemoradiotherapy,and patients are prone to relapse after surgery and the residue tumor will develop to higher-grade glioma.Although new treatment strategies have been proposed in the past decade and substantial progress has been made in the preclinical stage,the results in the clinical trial stage are not satisfactory.Complex tumor microenvironment,blood-brain barrier block to drug delivery,heterogeneity in the tumor,lack of effective tumor biomarkers and other factors bring difficulties to the early detection and treatment of gliomas.The TEAD protein family belongs to transcription factors,which include four family members:TEAD1,TEAD2,TEAD3,and TEAD4.TEAD transcription factors are involved in the progression of various cancers.But the role of TEAD3 in glioma has rarely been reported.ObjectiveTo detect the expression of TEAD3 transcription factor on glioma and the function of TEAD3 transcription factor on glioma proliferation、invasion、migration and angiogenesisMethod1.Comparing the expression of TEAD3 between glioma and normal tissues with TCGA、GTEX and GEO databases.And validating the result by qPCR.2.Performing survival analysis between TEAD3 high and low expression group gliomas combined with CGGA And TCGA database,and GO,KEGG,and GSEA enrichment analysis were used to preliminarily explore the function of TEAD3 on gliomas.3.Transfecting glioma cell lines U87 and A172 with TEAD3 overexpression lentivirus and siRNA to up-regulate or knock down the expression of TEAD3.Detecting the effects of TEAD3 on glioma invasion、migration、proliferation、angiogenesis by transwell invasion、migration assay、CCK8、plate colony formation assay and tube formation assay in vitro experiment.Detecting the effects of TEAD3 on glioma formation by subcutaneous tumor-bearing models in vivo experiment.4.The expression of hippo pathway target genes CTGF and CYR61 was detected by qPCR.And the expression of invasion and migration related proteins E-cadherin and MMP9 was detected by Western blot and cell immunofluorescence.Result1.TEAD3 expression on gliomas was higher than normal tissue,and high TEAD3 expression was associated with poor prognosis in gliomas.2.Extracellular matrix remodeling-related signatures and angiogenesis signature were significantly enriched in TEAD3 high expressed group of glioma.3.The results of transwell invasion and migration assay showed that knockdown of TEAD3 inhibited the migration and invasion of glioma cells U87 and A172,while the cell migration and invasion increased after overexpression of TEAD3.Western blot and immunofluorescence results showed that E-cadherin protein expression and fluorescence intensity increased,MMP9 protein expression and fluorescence intensity decreased in cells after TEAD3 knockdown.On the contrary,E-cadherin protein expression and fluorescence intensity decreased,and MMP9 protein expression and fluorescence intensity increased in cells after overexpression of TEAD3.4.The results of in tubule formation assay in vitro showed that the tubule formation ability of umbilical vein endothelial cells(HUVECs)was enhanced after co-culture with glioma cells with TEAD3 overexpression.The results of CCK8 cell proliferation and plate cloning assay showed that knockdown or overexpression of TEAD3 had no effect on the proliferation ability of glioma cells,and the hippo signaling pathway target genes CTGF and CYR61 were not significantly changed after overexpression of TEAD3.The results of subcutaneous tumor-bearing experiments showed that the growth rate of subcutaneous tumors in mice with TEAD3 overexpression was faster than that in control mice.Conclusion1.TEAD3 expression is elevated on gliomas and acts as a risk prognostic factor for glioma.2.TEAD3 can promote glioma invasion and migration by affecting E-cadherin and MMP9 expression.3.TEAD3 can promote glioma angiogenesis in vitro.4.TEAD3 does not affect tumor cell proliferation in vitro and may be functionally redundant with other TEAD family members.5.TEAD3 can promote glioma formation in vivo.