Screening Of Glioma Oncogene And The Molecular Mechanism By Which Oncogene YAP1 Promotes High Invasion Of Glioma Through STAT3-mediated Epithelial Mesenchymal Transformation

Objective: Glioma is one of the most common tumors of central nervous system,among which Glioblastoma(GBM)is the most aggressive and has a poor prognosis,and there is still no effective treatment.Epithelial-to-mesenchymal transition(EMT)is an important regulatory event of highly invasive glioma.Therefore,screening onco genes of glioblastoma and studying the molecular mechanism of oncogene-mediated EMT promoting the invasion of glioblastoma can provide theoretical basis for the clinical development of effective targeted therapy and improvement of prognosis.Methods: 1)Tandem mass tagging(TMT)was used to screen for differentially expressed proteins in 5pairs of GBM and non-tumor fresh tissues,and candidate markers were identified in combination with multiple bioinformatics analysis platforms and the characteristics of glioblastoma with high invasion and proliferation.2)Immunohistochemistry(IHC)is applied to determine the expression levels of YAP1(YAP1,p YAP1)and STAT3(STAT3,pSTAT3-S727,pSTAT3-Y705)in clinical glioma samples.Their relationship with clinicopathologi cal features and prognosis was analyzed,and the relationship between YAP1 and STAT3 was preliminarily evaluated.IDH1 gene mutation was detected by Sanger sequencing and co-deletion of 1P19 Q was detected by Fluorescence in situ hybridization(FISH),and of YAP1 and STAT3 expression in different molecular subtypes were analyzed.3)To establish stable YAP1 overexpressed strain,observe the effects of YAP1 and STAT3 on the biological functions of glioma cells,and preliminarily explore the interaction between YAP1 and STAT3 from cytology.The interaction between YAP1 and STAT3 was observed and verified by immunoprecipitation,immunofluorescence and nucleoplasmic protein isolation.The mechanism of YAP1 regulating EMT in glioma by STAT3 was studied at the cellular level.To observe the effect of YAP1 overexpression on temozolomide treatment.Results: 1)2387differentially expressed proteins were screened by proteomics,including 1206 differentially up-regulated proteins and 1181 differentially down-regulated proteins.In this study,combined with proteomics results,TCGA database and GBM high invasion characteristics,YES-associated protein 1(YAP1)and Signal transducers and Activators of transcription(STAT3)are candidate markers.2)The expression of YAP1 and PSTAT3-S727 was related to IDH1 mutation status,but the expression of YAP1 and STAT3 was not related to co-deletion status of 1P19 Q.The high expression of YAP1 and STAT3 in GBM is associated with poor prognosis of patients with glioma,there was a positive correlation between YAP1 and STAT3 in glioma samples.3)Cytological studies confirmed the interaction between YAP1 and STAT3.YAP1 promoted the proliferation,migration and invasion of glioma cells through STAT3.YAP1 regulates the EMT process of glioma through STAT3.High expression of YAP1 can reduce the therapeutic effect of temozolomide.Conclusion: YAP1 affects the biological function of glioma cells,mediates the EMT event of glioma and affects the therapeutic efficacy of temozolomide through STAT3.The YAP1-STAT3 signaling axis can be an important signaling pathway for targeted treatment of glioma and improvement of prognosis.