| BackgroundGlioma is the most common of the central nervous system tumors.its biological characteristics is high malignancy,strong invasiveness is strong,and high recurrence rate.Existing treatments are not ideal for the average prolonged survival of patients with glioma.Therefore,we need to conduct more in-depth research on the process of occurrence and development of gliomas at the molecular level,Seek more accurate diagnoses and more effective treatments.The tumor microenvironment is of great significance to the development of tumors,especially the immune-related tumor microenvironment,which plays a particularly important role in the molecular targeted therapy and immunotherapy of gliomas.In addition to being a programmed death of tumor cells,pyroptosis also has a certain impact on the tumor microenvironment.Therefore,the regulation of pyroptosis is also particularly important for the development of glioma.The shearing factor SRSF6 is expressed abnormally in a variety of tumors,and it has been found that gliomas are no exception,and it is likely to be involved in the regulation of glioma cell scorch death,then affects the occurrence and development of gliomas.At the same time,ZNf263 was also found as a transcription factor of SRSF6,which could regulate SRSF6 protein expression,and then participated in the regulation of glioma cell pyroptosis.SRSF6-regulated glioma cell pyroptosis has a certain impact on the tumor microenvironment,in which the granzyme B,an important executive molecule produced and secreted by CD8+ T cells,has undergone significant changes,further promoting changes in the biological behavior of glioma.SRSF6 regulates the occurrence of glioma pyroptosis,and the impact on the tumor microenvironment further promotes glioma cell death as a feasible research scheme,and there are no studies being reported in relevant fields.ObjectiveThe objective of this study was to research the effect of ZNF263 on the transcriptional regulation of SRSF6,the effect of SRSF6 on glioma cell pyroptosis,and the interaction of glioma cells with the tumor microenvironment by using glioma cell lines and animal glioma models,and to explore its molecular mechanism.MethorsIn this study,the differences in the expression of SRSF6 gene in normal tissues and gliomas were predicted by TCGA database and GTEx database;the differences in the expression of SRSF6 genes and proteins in glioma tissues and cell lines were analyzed by quantitative PCR and western blot;to detect the differences in the expression of SRSF6 in individual grades of gliomas by immunohistochemistry;The clinical information of patients with glioma disease who admitted to our department in the past was collected,and the relationship between SRSF6 and patient survival was examined through immunohistochemistry analysis of SRSF6 protein expression score and clinical multivariate correlation in patients with glioma.Construct a stable SRSF6 gene interfered glioma cell line,observe the change of glioma cell activity after interfered with SRSF6 expression by CCK8 experiment,and observe the relationship between the expression and secretion of important molecules related to pyroptosis and the expression of SRSF6 protein when glioma cell pyroptosis occurs.The effect of SRSF6 on the development of tumorigenesis by regulating pyroptosis was observed by the nude mouse subcutaneous tumor model.The mechanism of ZNF263 regulates SRSF6 protein expression was verified by double luciferase gene reporting experiment and chromatin immunoprecipitation experiment,and ZNF263 participated in regulating the occurrence of glioma cell pyroptosis and promote the development of glioma by regulating SRSF6.Construct mouse intracranial glioma model,and the relationship between the changes of important immune-related molecules in the tumor microenvironment and the expression of SRSF6 was observed by immunohistochemistry,and the expression changes of pyroptosis-related molecules in mouse glioma cell lines were observed by western blot.The effect of granzyme B on apoptosis of glioma cell was observed by flow cytometry.Apply statistical analysis software to statistically analyze relevant data to determine whether it is statistically significant.Result1.SRSF6 gene and protein are highly expressed in glioma tissue relative to tissue;the expression of SRSF6 is positively correlated with the pathological grade of glioma tissue,that is,the higher the glioma level,the higher the expression of SRSF6 protein;the survival of glioma patients with high expression of SRSF6 is significantly lower.2.SRSF6 can inhibit the pyroptosis of glioma cell,and can significantly inhibit tumor growth after knocking down SRSF6 in nude mouse subcutaneous tumor model.3.ZNF263 can enhance the transcription of SRSF6 by binding to the promoter region fragment of SRSF6 gene,thereby promoting the expression of SRSF6 protein;ZNF263 can inhibit glioma cell coke death by promoting SRSF6 protein expression,and tumor growth can be significantly inhibited after knocking down ZNF263 in nude mouse subcutaneous tumor model.4.In the mouse intracranial glioma model,pyroptosis regulated by SRSF6 can change the immune response related to the tumor microenvironment,and knocking down SRSF6 in the mouse glioma model,CD8+ T cells are increased and the production of granzyme B is increased;granzyme B can promote apoptosis of human and mouse glioma cells.ConclusionSRSF6 is highly expressed in glioma;SRSF6 expression is inversely correlated with the survival of patients with glioma,which can be used as a potential prognostic evaluation criterion;in glioma cells and glioma animal subcutaneous tumor models,SRSF6 can inhibit the occurrence of glioma cell pyroptosis,ZNF263 can enhance the expression of SRSF6 protein,and inhibit pyroptosis.At the same time,SRSF6 can inhibit the generation of granzyme B by inhibiting the pyroptosis of glioma cells and thereby inhibiting the generation of granzyme B by CD8+ T cells in the glioma microenvironment,reducing the apoptosis of glioma cells caused by granzyme B,and indirectly promoting tumor cell proliferation and tumor development. |
PDF Full Text Request