Application of TRIM24 in the diagnosis of glioma
A glioma, p-egfry1172 technology, applied in the application field of TRIM24 in the diagnosis of glioma, can solve the problems of unfavorable comprehensive treatment and improvement of curative effect, inaccurate pathological diagnosis, lack of pathological basis for treatment, etc. Improve the quality of life, increase survival, the effect of targeted and effective medication
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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 form, 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 a...
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 ) were 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 3...
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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