PARD6A Regulate The Proliferation And Stemness Maintenance Of Malignant Glioma

Background: Glioma is the most common and most lethal tumor in the brain.According to WHO classification,gliomas are classified into four grades.Glioblastoma is grade IV.Glioblastoma is the most common malignant tumor of the central nervous system,with a5-year survival rate of only 5%.The current conventional treatment for glioma,a combination of surgery,radiation and chemotherapy with temozolomide,has improved the overall survival of glioma patients from an average of 16 months to 20.9 months.However,this therapy still cannot solve the problems of poor prognosis and recurrence in patients with glioma.One of the reasons for the poor prognosis of patients with glioblastoma is that under standard treatment,glioblastoma remains a population of stem cells that can evade radiation and chemotherapy and cause recurrence after treatment.Glioma stem cells are a group of cells with the ability to self-renew,promote migration,invasion and tumorigenesis.Therefore,targeting glioma stem cells has become a target for glioblastoma therapy.Objective: In this study,we explored the mechanism of PARD6 A synergistic with SOX2 in promoting the dry maintenance and tumor-forming ability of glioma,so as to screen out the specific short peptide P1 targeting the complex,and provide theoretical basis and experimental basis for further optimizing the treatment plan and current situation of glioma.Methods: 1.GSEA was used to predict the effect of PARD6 a on dry maintenance of glioma and to predict the protein molecules forming complex with PARD6 a by stem cell limit dilution test,pelletogenesis test,Western blotting and in vivo animal imaging.2.The complex formed by PARD6 a and SOX2 was verified by immunofluorescence,co-IP,prokaryotic expression and GST-pull down experiments,and the specific binding region of the complex was determined.3.Phage technology,CCK8,GST-pull down and in vivo imaging of small animals were used to investigate the effects of specific short peptide P1 on dry maintenance and tumorigenesis ability of malignant glioma.Results: 1.GSEA,stem cell limit dilution test,pellet-forming test,Western blot and in vivo imaging of small animals showed that overexpression of PARD6 a significantly promoted the dry maintenance and tumor-forming ability of malignant glioma.2.According to GEPIA website prediction,Western blotting,stem cell limit dilution assay and pellet-forming assay,PARD6 a and SOX2 were positively regulated,and SOX2 significantly promoted the dry maintenance of glioma.3.Through immunofluorescence,co-IP,prokaryotic expression and GST-pull down experiments,we can see that there is an interaction between PARD6 a and SOX2,and the interaction is direct.The specific binding region of PARD6 A and SOX2 was determined.4.Phage display technology was used to screen out three specific short peptides.CCK8,GST-pull down,stem cell limit dilution test and pellet forming test showed that the specific short peptide P1 significantly inhibited the dry maintenance and tumorigenesis ability of malignant glioma.Conclusion: The results of this study showed that: 1.PARD6 A and SOX2 interact to form a complex,and this complex promotes the dry maintenance of malignant glioma;Overexpression of PARD6A/SOX2 complex promotes tumorigenesis of malignant glioma.2.The specific short peptide P1 screened for PARD6A-1 significantly inhibited the proliferation,dry maintenance and tumorigenesis of malignant glioma.Based on the research of this topic,PARD6A/SOX2 complex plays an important role in the pathogenesis of glioma.The regulatory relationship of this complex and its biological characteristics are described,which provides a new idea for elucidating the pathogenesis of glioma and also provides a basis for elucidating the mechanism of glioma dry maintenance.And to find out the target of glioma stem cell therapy provides a new theoretical and experimental basis.