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A glioma prognostic marker and its application

A technology for glioma and prognosis, applied in biochemical equipment and methods, microbiological measurement/testing, DNA/RNA fragments, etc., can solve problems such as poor survival rate

Active Publication Date: 2021-07-02
XUZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vast majority of patients with glioma are in the middle and advanced stages, and the survival rate after surgery is not optimistic

Method used

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  • A glioma prognostic marker and its application
  • A glioma prognostic marker and its application
  • A glioma prognostic marker and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Glioma Prognostic Diagnostic Kit:

[0020] ① RNA stabilization solution 50ml

[0021] ② TRIzol (Invitrogen) 50ml

[0022] ③ Chloroform (Sinopharm Chemical Reagent Co., Ltd.) 100ml

[0023] ④ Isopropanol 100ml

[0024] ⑤Nuclease-free water 10ml

[0025] ⑥4×gDNA wiper Mix (Nanjing Nuoweizan Biotechnology Co., Ltd.) 400ul

[0026] ⑦5×HiScriptⅡSuperMixⅡ(Nanjing Nuoweizan Biotechnology Co., Ltd.) 400ul

[0027] ⑧2×AceQ qPCR SYBR Green Master Mix (Nanjing Nuoweizan Biotechnology Co., Ltd.) 400ul

[0028] ⑨10μM lnc RNA TCONS_00020456 real-time fluorescence quantitative PCR specific primer 30μl,

[0029] lnc RNA TCONS_00020456 forward primer: 5'-CTCAGGTGGTGCCATTCTC-3',

[0030] lnc RNA TCONS_00020456 reverse primer: 5'-GACCTTGTCCTGCTCTTCATT-3', synthesized by Shanghai Sangon Bioengineering Co., Ltd.;

[0031] ⑩10μM HsActin specific primer 30μl

[0032] Forward primer: 5'-CGGGAAATCGTGCGTGAC-3',

[0033] Reverse primer: 5'-GCCCAGGAAGGAAGGCT-3'.

Embodiment 2

[0035] From the glioma samples obtained from the Affiliated Hospital of Xuzhou Medical University from 2016 to 2018, tissue RNA was extracted, and low-grade and paracancerous tissues were used as controls to compare the levels of TCONS_00020456 in different grades of glioma tissues:

[0036] ①Extract tissue RNA: Take an appropriate amount of sample and bake it at 180°C for 6-8 hours, add liquid nitrogen to the mortar to grind the sample, grind it to powder, add 1ml TRIzol mortar sample in the mortar, grind it into liquid, and move to Add 200 μl of chloroform to the tube, shake it by hand for 15-30 seconds, place it on ice for 15 minutes, and centrifuge at 12000 rpm at 4°C for 15 minutes; carefully take the upper aqueous phase into a new tube, add 0.5ml of pre-cooled isopropanol and mix well. Let stand on ice for 20min, centrifuge at 12000rPm at 4°C for 10min; discard the supernatant, add 1-2ml of ethanol diluted with 75% DEPC water and mix well, centrifuge at 7500rPm at 4°C for...

Embodiment 3

[0055] Patients diagnosed with glioma in the Affiliated Hospital of Xuzhou Medical University from 2006 to 2009 were surgically resected. After surgery, 148 cases of tissue were fabricated to obtain glioma tissue chip arrays, which were reserved for research use. The study used in situ hybridization combined with immunohistochemistry to stain the tissue chip with TCONS_00020456, and then scored the results one by one (according to the proportion of positive cells and the color depth), and then made high-grade glioma tissues and WHO grade I tumors. Tissue ROC curve, see figure 2 , figure 2It is the ROC curve of the markers of the present invention for high-grade glioma and WHO grade I tumor tissue. It can be seen that the AUC is 0.89 (CI=0.82-0.97, sensitivity=79%, specificity=89%), This shows that TCONS_00020456 has a high diagnostic value for malignant glioma.

[0056] The results of 148 cases of microarrays in in situ hybridization were processed as follows: In order to ...

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Abstract

The invention relates to a glioma prognosis marker and belongs to the field of biotechnology. The marker is lnc RNA TCONS_00020456, the sequence of which is shown in SEQ ID NO.1. The sequences of the specific primers used to detect the expression levels of the above markers are: F: 5'-CTCAGGTGGTGCCATTCTC-3', R: 5'-GACCTTGTCCTGCTCTTCATT-3'. The present invention uses a qRT-PCR detection method to detect the expression level of lnc RNA TCONS_00020456 in glioma tissue sources of patients, and studies have confirmed that patients with higher expression levels in glioma have higher postoperative survival rates. Therefore, lnc RNA TCONS_00020456 can be used as a prognostic marker of glioma for prognostic analysis of glioma patients.

Description

technical field [0001] The invention relates to a glioma prognosis marker and belongs to the field of biotechnology. Background technique [0002] Tumors derived from the neuroepithelium are collectively referred to as gliomas, which account for 40% to 50% of brain tumors. They are the most common intracranial malignant tumors, with an annual incidence of 3 to 8 per 100,000 population. , high recurrence rate, high mortality rate and low cure rate. [0003] At present, the treatment of glioma is mainly through surgical resection, supplemented by radiotherapy and chemotherapy. However, postoperative patients have a high recurrence rate. One of the main factors that make glioma difficult to cure is the aggressive growth of glioma itself, the boundary between tumor cells and normal brain tissue is very blurred, and it is difficult to completely remove it by surgery. In addition, the rapid proliferation of glioma cells leads to The formation of the local tumor hypoxic microenv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/178
Inventor 唐传喜王月高殿帅王洁张磊
Owner XUZHOU MEDICAL UNIV
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