The Research Of RSRP1 Promoting Malignant Progression Of Glioma And Its Molecular Mechanism

Glioma is the most common primary tumor in the central nervous system,and more than half of them are the most malignant glioblastomas(GBM),even if the current most standard treatment is adopted:with the maximum safety range surgical resection and supplemented with standard concurrent chemoradiotherapy,the patient’s prognosis was still very poor,with a median survival of only 14.6 months.The main cause of poor prognosis of glioma patients is that glioma has a malignant phenotype with strong proliferation ability,genetic background heterogeneity,and resistance to radiotherapy and chemotherapy.Therefore,the search for new therapeutic methods that can effectively against the malignant progression of gliomas,prolong the survival of patients with glioma,and improve the quality of life of patients is the focus of glioma research.Our previous research focused on the study of tumor cells outside the glioma tumor entity and found that there are glioma cells in the peripheral edema zone in glioma patients,and their biological characteristics is different from cells in tumor entity("C cells").We named this kind of cells as "Q cells".Our laboratory believes that these tumor cells present in difficult-to-remove areas are the culprits for repeated recurrences and the difficulties of eradication of glioma.Research on Q cells is expected to truly solve the world problem of refractory glioma.With the continuous development of research techniques in molecular biology,people’s understanding of cancer gradually deepens.The WHO revised 2016 CNS firstly integrates molecular subtyping classification into the current histological classification of gliomas,breaking the tradition of histological classification in the past 100 years and understanding gliomas.This alteration signified that our knowledges of glioma has reached an unprecedented depth.In the past,the molecular phenotypic characteristics of gliomas were mostly related to genes,providing multiple molecular pathological indicators for the diagnosis of gliomas,such as IDH1 mutations,chromosome 1p/19q co-deletions,TERT gene mutations,and p53 gene mutations.There are also many studies that have turned their attention to the level of transcription and explored the specific expression of genes in gliomas.Alternative splicing is an important process before the gene is translated into a biologically functional protein after being transcripted into a precursor mRNA,and there is no shortage of studies on the alternative splicing of gliomas,confirming that the same gene is under the control of splicing factors.Different splice isoforms play a different biological role in gliomas.The regulation of alternative splicing affects the structure of the protein that ultimately functions,and the study of splicing isomers can more directly and truly reflect the significance of the molecule in tumors,and has important prospects in molecular research of gliomas.Our laboratory used transcriptome sequencing to discover the differences between the expression patterns of Q cells and C cells.We tried to explain the biological behavior of Q cells through in-depth exploration of differentially expressed factors,providing new ideas for the treatment of glioma.RSRP1(arginine/serine-rich protein 1)is a novel molecule high expressed in Q cells.In this study,bioinformatics analysis and molecular biological experiments confirmed that it regulates alternative splicing to perform biological function in glioma.At the same time,after analyzing the clinical data of the glioma public database platform and the neurosurgery glioma specimens of the Nanfang Hospital,the clinical significance of RSRP1 in gliomas was clarified.And the role RSRP1 play in the malignant progression of gliomas was clarified through in vitro and in vivo experiments.Section 1 The biological function of RSRP1 performed in malignant progression of glioma and its clinical significancePurpose:To investigate the clinical significance of the novel molecule RSRP1 in glioma and its participation in biological functions.Methods:Download the mRNA expression profile data and clinical data of glioma public data platform(TCGA,GEO),and use bioinformatics analysis to analyze the correlation between relative expression of RSRP1 mRNA in the glioma samples and the prognosis of the patients.Immunohistochemical method was used to detect the relative expression of RSRP1 in clinical specimens of gliomas.The clinical data was used to analyze the correlation between the expression of RSRP1 protein in gliomas and the prognosis of patients.The expression of RSRP1 was interfered in the glioma cell line by silencing,and the biological function(proliferation,apoptosis,invasion,and migration)of the post-interference glioma cells was examined in vitro and in vivo.Results:The results of clinical data analysis showed that high expression of RSRP1 was negatively correlated with the prognosis of patients with glioma;after interfering with the expression of RSRP1,the apoptosis of glioma cells increased and the invasion and migration ability decreased.Conclusion:RSRP1 is a good indicator for predicting the prognosis of patients with glioma.RSRP1 high expression indicates poor prognosis;RSRP1 participates in apoptosis resistance and promotes invasion and migration in malignant progression of glioma.Section 2 RSRP1 promotes the malignant progression of glioma by modulating alternative splicingPurpose:To explore the molecular mechanism of RSRP1 promoting the malignant progression of glioma.Methods:Using bioinformatics analysis to predict the molecular biological processes involved in RSRP1,mainly through the analysis of RSRP1 domain,interacting proteins,co-expressed genes in three parts to predict.The specific mechanism of RSRP1 involvement in glioma function was verified by transcriptome sequencing and RTPCR.Yeast two-hybrid and protein co-immunoprecipitation experiments were performed to verify the interaction protein of RSRP1.Results:Bioinformatics analysis suggests that RSRP1 may be associated with alternative splicing regulation of genes in gliomas.The results of transcriptome sequencing suggested that RSRP1 may regulate the splicing of a large number of downstream tumor-associated genes.RTPCR further confirmed the sequencing results.Co-immunoprecipitation experiments confirmed that RSRP1 interacts with alternative splicing-related proteins.Conclusion:RSRP1 participates in the biological process of malignant progression of glioma by modulating the splicing form of tumor-associated genes in glioma cells.