Application of FAM13A gene transcripts in evaluating clinical prognosis of glioblastoma patients
A glioblastoma, FAM13A-004 technology, applied in the field of medicine, can solve problems such as poor treatment effect, achieve the effect of optimizing treatment plan and improving overall survival
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Embodiment 1
[0041] Example 1 RBMX expression level to evaluate glioblastoma subtype and prognosis
[0042] 1. Screening the transcriptional regulators related to the prognosis of glioblastoma through the database
[0043] The GBM transcriptome typing system of TCGA (The Cancer Genome Atlas) is currently the most commonly used typing system, which includes the preneural type (Proneural) and the mesenchymal type (Mesenchymal). The two subtypes are closely related and often involve EMT progress in GBM. In the present invention, through the subtype annotation of more than 1,000 GBM samples from the TCGA database and the GSE16011 database, 404 transcription factor gene expression differences were screened between Proneural GBM and Mesenchymal GBM, and then combined with TCGA-U133A, Agilent, HiSeq and GSE16011 multi-platform results and clinical prognosis data found that RBMX was lowly expressed in Mesenchymal GBM and highly expressed in Proneural GBM, and the survival time of glioblastoma pat...
Embodiment 2
[0067] Example 2 Correlation detection between RBMX expression and EMT markers
[0068] RBMX is low expressed in Mesenchymal glioblastoma and highly expressed in Proneural glioblastoma. RBMX, as a transcription factor, is involved in the process of EMT. The expressions of EMT-related markers E-cadherin, Claudin1, N-cadherin, Slug and β-catenin in knockdown and overexpression RBMX cell lines were detected by Western blot (WB), immunofluorescence and immunohistochemistry .
[0069] 1. Immunofluorescence technique
[0070] (1) Divide 5×10 3 Spread the cells evenly on the cell slides of the 24-well plate overnight to ensure the adherent growth of the cells;
[0071] (2) Fix the cells with 4% paraformaldehyde, permeate the membrane with PBS solution containing 0.5% Triton, and seal the cell slide with 5% fetal bovine serum (BSA);
[0072] (3) Add E-cadherin, Claudin1, N-cadherin, Slug and β-catenin specific antibodies (primary antibodies) to the cell slides, add 200 μL to each...
Embodiment 3
[0087] Example 3 Using qPCR technology and agarose gel electrophoresis technology to detect the expression levels of transcripts FAM13A-004 and FAM13A-001
[0088] RBMX-encoded RNA-binding proteins play diverse regulatory roles in pre- and post-transcriptional processes, and are involved in tissue-specific regulation of gene transcription and selective transcription of pre-mRNAs. By analyzing the TCGA transcript sequencing data, we found that the expression of RBMX was correlated with the expression level of more than 7000 transcripts (r>0.3 or Figure 4 a. The FAM13A-004 transcript contains a RAS homologous GTPase-activating protein (RhoGAP, Ras homologous GTPase-activating protein) domain, which affects GTPase activity, down-regulates the RhoA signaling pathway, inhibits the progression of EMT, and inhibits the malignant proliferation of GBM. The FAM13A-001 transcript contains two coiled coil domains (CCDs), which are involved in protein-protein interactions. By binding to t...
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