The Effect Of Transcription Factor RelB On The Growth And Invasion Of Glioma And Its Mechanism

Part 1 The biological function of transcription factor Rel B in gliomaObjective:To investigate the expression of transcription factor Rel B in different grades of gliomas.To observe the changes in the expression of the transcription factor Rel B in different subtypes of glioma.To investigate the relationship between transcription factor Rel B and prognosis of glioma.To discuss the changes in the growth ability of the glioma after down-regulation of transcription factor Rel B.To study the effect of inhibiting transcription factor Rel B on the invasion and metastasis of glioma cells.Method: Choosing 325 cases of gliomas in CGGA database tissue samples as research object,on the gene sequencing,andomly divided into three groups of malignant degree,studied the difference in the expression of transcription factor Rel B in gliomas with different malignant degrees.Immunohistochemical method was used to detect the expression of Rel B protein in 32 cases of gliomas in our hospital.TCGA database was selected to analyze the expression of transcription factor Rel B in glioma cells with different molecular subtypes.The CGGA database of 325 glioma patients according to the degree of malignancy was divided into low grade gliomas,anaplastic gliomas,glioblastoma.To analyze the effect of Rel B expression on prognosis of patients with different grade gliomas.Transfection of Rel B inhibitor with cationic liposome in glioma cell line.The expression of Rel B in glioma cell line treated with Rel B inhibitor was detected by Western blot(Western-blot),and the transfection efficiency was evaluated.CCK8 method was used to observe the effect of Rel B on the growth of glioma cells.Flow cytometry was select to detect the changes of cell cycle after treatment with Rel B inhibitor.Detection of cell migrationin glioma cell line by scratch test.Transwell assay was choosed to detect the invasion and migration of glioma cells.Results: In the CGGA database,with increasing the degree of malignancy of gliomas,the expression of Rel B increased;confirmed in clinical glioma patients,immunohistochemistry results showed that with the increased level of glioma,the expression level of Rel B protein increased gradually.The results of the analysis of glioma data in TCGA database showed that the Rel B expression was different in different subtypes of glioma tissue.Compared with the control group,the high expression of Rel B was associated with poor prognosis in patients with Anaplastic glioma and glioblastoma.After transfection with Rel B inhibitor,Western blot showed that the expression of Rel B protein decreased significantly.CCK8 showed that the decrease of Rel B expression inhibited the proliferation of tumor cells.With the decrease of Rel B,cell cycle arrest in G0-G1 phase.The invasion and migration ability of glioma cells was significantly decreased.Conclusion: Our results showed that high expression of Rel B promotes the growth,invasion and migration of glioma cells,which is closely related to the grade and subtypes of gliomas,suggesting a poor prognosis.Part 2 Molecular mechanism of transcription factor Rel B in gliomaObjective: To investigate the role of transcription factor Rel B in the development and progression of glioma.Searching downstream target gene of transcription factor Rel B.To verify the regulation of transcription factor Rel B on target gene MMPs.To validate the effect of transcription factor Rel B on the development and progression of glioma through the mechanism of the expression of target gene MMPs.Method:Choosing 325 cases of gliomas in CGGA database,the application of Gene Ontology Analysis determine transcription factor Rel B protein domain function.Take the Pearson Correlation analysis screening gene sequence data of glioma specimens obtained target gene.Glioma cell line U87 and LN229 were selected,in which cationic liposome was used to transfect Rel B inhibitor.After transfection with Rel B inhibitor,the expression of transcription factor Rel B was decreased by Western blot,and the expression level of MMP2 and MMP9 in the matrix metalloproteinases decreased significantly.Results: GO analysis showed that the function of transcription factor Rel B protein domain was closely related to the cell cycle,invasion and metastasis.Pearson Correlation Analysis was used to verify the RNA sequence data of glioma specimens and the target gene was MMPs.After transfection with Rel B inhibitor,Western blot showed that the expression of Rel B protein decreased significantly.Conclusion: The transcription factor Rel B is closely related to the growth,invasion and migration of glioma cells through the matrix metalloproteinases family(MMPs),especially MMP2 and MMP9.