Functional Study Of DUSP10 In Glioma And Anti-glioma Effects Of Dipine Drugs

Objective: Glioma is a malignant tumor originating from glial cells in the central nervous system.Due to its biological characteristics of rapid proliferation,invasion,infiltration and growth,anti-apoptosis,angiogenesis mimicry and other biological characteristics,glioma seriously endangers the life safety of patients,and brings a huge burden to family members and social medical security system.At present,surgery combined with chemoradiotherapy and other means are commonly used in clinical comprehensive treatment,but its five-year survival rate is extremely low,only better than pancreatic cancer and lung cancer among all tumors,ranking the third from the bottom.Therefore,from the clinical perspective,the author mined the sequencing data of clinical specimens of glioma patients through bioinformatics to deeply explore the biological principle of the occurrence and development of glioma,screen out the proto-oncogenes related to the malignant process of glioma,and explore the therapeutic drugs that can inhibit the proto-oncogenes on the market with the help of the high-throughput drug screening platform of Chinese Academy of Sciences.The therapeutic effect and its underlying mechanism were quantified through experiments.It is hoped that the author’s study can provide a certain laboratory basis for the follow-up exploration of glioma prognosis and therapeutic targets.Methods: In order to explore the genetic background of glioma,the author started from the bioinformatics,and adopted The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)to explore the genetic background of glioma.Combined with the Chinese Glioma Genome Atlas(CGGA)and other databases independently developed by China,the clinical prognostic analysis of genes affecting the prognosis of glioma was carried out.A number of pathogenic genes which may have certain functions in the occurrence and development of glioma were enriched.Finally,we found that DUSP10 may profoundly affect the prognosis and survival of glioma patients.To this end,we conducted wet experiments including DNA methylation,immune cell infiltration,signal pathway enrichment,proliferation,apoptosis,and motor invasion,and revealed the phenotype and possible molecular mechanism of DUSP10 affecting the poor prognosis of glioma patients.At the same time,the author conducted a secondary screening of the antitumor drugs previously screened in the laboratory by WB assay,and found that two calcium channel blockers,amlodipine and felodipine,could significantly reduce the expression of DUSP10.Meanwhile,through IC50,EDU,immunofluorescence,Annexin V-FITC and other tests,we quantified the killing efficiency of two calcium channel blockers against glioma cell lines and stem cell lines.Finally,ROS experiment,Jc-1 experiment and Western blotting experiment were used to explore the internal mechanism of drug killing glioma cell lines,and our conclusions were further verified by rescue experiment.Results: By TCGA pancarcinoma analysis,we found that DUSP10 was highly expressed in a variety of tumors,including GBM and LGG,and deeply affected the prognosis of patients with a variety of tumors.Through public database analysis,we found that DUSP10 expression level was significantly negatively correlated with1p/19 q codeletion,IDH1 mutation and other favorable mutations,and positively correlated with WHO grade of glioma.Further COX analysis and nomogram showed DUSP10 as an independent prognostic indicator.It had a good correlation with overall survival(OS),Progression-free Survival(PFS)and disease-specific Survival(DSS).In addition,the methylation degree of DUSP10 promoter region in glioma patients was significantly lower than that in normal control group,suggesting that DNA methylation may be an important cause of DUSP10 overexpression in glioma patients.In terms of mechanism,through GSEA,GO and KEGG analysis,we found that DUSP10 was associated with T cell proliferation and activation and enriched in JAK-STAT,cell cycle and NF-kappa B signaling pathways.Wet experiment results showed that the proliferation and migration of glioma cell lines T98 G and A172 were significantly reduced after DUSP10 was knocked down,while apoptotic cells were increased,which indicated that DUSP10 played an important role in the development of glioma.On the other hand,through the database prediction and drug screening data,we quantified the killing effect of drugs on glioma,and found that amlodipine could significantly inhibit the proliferation ability and induce apoptosis of glioma cell lines,and the IC50 value was lower than that of Temozolomide(TMZ),the clinical first-line chemotherapy drug.Further pathway experiments revealed that the two drugs initiate autophagy by inhibiting the PI3K-AKT pathway,leading to cell death.Conclusion: DUSP10 is a proto-oncogene,which promotes the proliferation and movement of gliomas and inhibits cell apoptosis in the development and progression of gliomas.DUSP10 can be used as an independent prognostic factor to profoundly affect patients’ clinical survival indicators such as OS,PFS and DSS.Dusp10 is expected to become a target for clinical prognosis and treatment of glioma patients.In terms of drug screening,through drug simulation combination,we found that DUSP10 may be combined with antihypertensive drugs felodipine and amlodipine.Meanwhile,compared with TMZ,felodipine and amlodipine can kill glioma cell lines and glioma stem cells more efficiently via PI3K-AKT axis.Our findings also provide laboratory evidence for the use of the two drugs in the treatment of glioma.