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Application of TRIM24 in glioma diagnosis

A glioma and gene technology, which can be applied to the determination/inspection of medical preparations containing active ingredients, antitumor drugs, microorganisms, etc., can solve the disadvantages of comprehensive treatment and improvement of curative effect, inaccurate pathological diagnosis, and lack of treatment Pathological basis and other issues, to achieve targeted and effective medication, improve survival, improve the effect of quality of life

Active Publication Date: 2018-06-26
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in my country, especially in some small and medium-sized hospitals, due to the lack of skilled neuropathologists, the pathological diagnosis after surgery is not accurate enough, and the WHO classification has not been adopted in some areas, so that the follow-up treatment of patients after surgery lacks reliable pathological basis
This is very detrimental to the comprehensive treatment of patients and the improvement of curative effect, and also makes it more difficult to evaluate the clinical curative effect

Method used

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  • Application of TRIM24 in glioma diagnosis
  • Application of TRIM24 in glioma diagnosis
  • Application of TRIM24 in glioma diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Increased Genomic Expression Level of TRIM24 Gene in Clinical Glioma Tumor Specimens

[0041] (1) Bioinformatics analysis 1:

[0042] Open the oncomine database (https: / / www.oncomine.org / ), enter TRIM24 in the search bar, select brain and CNS cancer in the Disease Summary for TRIM24 table, and select cancer VS.Normal Analysis in the primary filters on the left , select the database containing Glioblastoma vs.Normal in the results on the right to view the results.

[0043] The result is as figure 1 a shows: from TCGA data analysis shows that the average expression value of TRIM24mRNA in 10 cases of normal brain tissue is (log 2 )2.403, while the average expression value in 542 glioblastoma patients was 3.678(log 2 ), which is 2.503 times that of normal brain tissue. The expression level of TRIM24 in glioblastoma patients is significantly higher than that of normal brain tissue, and there is a statistical difference (p=1.17E-8).

[0044] (2) Bioinformatics ...

Embodiment 2

[0046] Example 2: Exogenous EGFRvlII significantly increases the expression level of TRIM24 in glioma U87 and LN229 cells

[0047] EGFRvIII was infected into glioma U87 and LN229 cells by lentivirus, then RNA was extracted, and the expression of TRIM24 was detected by qRT-PCR.

[0048] 2. 1293T cell packaging lentiviral particles:

[0049] 293T cells (purchased from ATCC, ATCC number CRL-3216 TM ) cultured in DMEM medium: containing 10% fetal bovine serum (Sigma, 12006C), 1% penicillin and streptomycin (Gibco, 15140-122) and DMEM medium (Gibco, 10564011), at 37 ° C, 5% CO2 without Bacteria incubator (thermo) culture.

[0050] On the first day, the 293T cells (the number of cells is about 5 × 10 6 ), aspirate the culture medium. Gently wash the cells with 2 mL DPBS (Invitrogen, 14190250) to remove residual medium, gently suck off the DPBS (be careful not to blow away the 293T cells), and repeat 3 times. After adding 1ml of 0.25% trypsin (Gibco, 25200-072) to digest at 37°C...

Embodiment 3

[0071] Example 3: In glioma-derived cancer cell lines, EGF stimulates cell proliferation and survival requires TRIM24 mediation

[0072] (1) Using the fugen-6 kit (purchased from Roche, 04709705001) according to the experimental steps of the instructions, two shRNA viral plasmids of TRIM24 (shT24-1 viral plasmid, Jikai Gene, 22927, shT24-2 viral plasmid, Jikai Gene, 22930) and shRNA control virus plasmid (shC virus plasmid, Jikai gene, CONO77) and psPAX2 (packaging plasmid, Addgene, Plasmid 12260), pMT24G (coating plasmid, Addgene, Plasmid 12259) formed virus infection to embodiment 2 In the obtained LN229 / EGFRvIII and U87 / EGFRvIII cells (LN229, U87 cells were purchased from ATCC, ATCC numbers are CRL-2611 TM and HTB-14 TM ).

[0073] 3. 1293T cell packaging lentiviral particles:

[0074] 293T cells (purchased from ATCC, ATCC number is CRL-3216 TM ) cultured in DMEM medium: containing 10% fetal bovine serum (Sigma, 12006C), 1% penicillin and streptomycin (Gibco, 15140-122)...

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Abstract

The invention discloses application of EGFR and TRIM24 coexpression in glioma diagnosis and provides a kit used for diagnosing the malignant degree of the glioma or predicating the lifetime of a glioma patient. The kit is characterized by comprising an agent which is used for detecting the expression level of the TRIM24 gene and / or protein, or the EGFR gene and / or protein and the TRIM24 gene and / or protein, and the TRIM24 gene and / or protein and the EGFRVIII gene and / or protein. The invention provides a new molecular marker of the glioma, the invention further designs a molecular marker kit which aims at the molecular marker and is used for judging the malignant degree of the glioma or predicating the lifetime of the glioma patient, and it is hopeful that a novel glioma therapeutic drug specifically aiming at a p-EGFRY1172 / TRIM24 signal circuit will be developed in future and the therapeutic effect of the therapeutic drug is verified.

Description

technical field [0001] The invention relates to the application of EGFR-TRIM24 co-expression in diagnosis and treatment of neuroglioma. Background technique [0002] Glioma is the most common primary intracranial tumor and mainly includes 4 pathological types: astrocytoma, oligodendroglioma, ependymoma, and mixed glioma. According to statistics from the American Brain Tumor Registry, malignant gliomas account for about 70% of primary malignant brain tumors. Among malignant gliomas, anaplastic astrocytoma (AA, WHO grade III) and glioblastoma multiforme (GBM, WHO grade IV) are the most common, with GBM accounting for approximately 50% of all gliomas . Currently, the diagnosis of glioma mainly relies on CT and MRI. Some new MRI, such as DTI, DWI, PWI, MRS, fMRI are helpful to improve the diagnosis and prognosis. PET and SPECT are helpful to distinguish tumor recurrence from radiation necrosis. Ultimately, however, tumor resection or biopsy is still required to confirm the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/00G01N33/68G01N33/574
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/156C12Q2600/158G01N33/57407G01N33/6863G01N33/6872
Inventor 冯海忠吕德官张伟伟
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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