Application of lncRNA-NEAT1 as molecular pathological diagnosis marker in preparation of glioma diagnosis reagent

A technology of pathological diagnosis and diagnostic reagents, which is applied in the field of medicine, can solve the problems of high price, poor sensitivity, and long experiment cycle, and achieve the effect of low cost, high sensitivity, and time period

Inactive Publication Date: 2018-03-09
HARBIN MEDICAL UNIVERSITY
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  • Application Information

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Problems solved by technology

[0006] The second object of the present invention is to establish a glioma diagnostic kit and its diagnostic method, to overcome the traditional in situ hybridization technology for detecting glioma-related lncRNAs which is expensive, long experiment cycle and poor sensitivity. Disadvantages to meet the needs of large-scale experiments and clinical applications

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  • Application of lncRNA-NEAT1 as molecular pathological diagnosis marker in preparation of glioma diagnosis reagent
  • Application of lncRNA-NEAT1 as molecular pathological diagnosis marker in preparation of glioma diagnosis reagent
  • Application of lncRNA-NEAT1 as molecular pathological diagnosis marker in preparation of glioma diagnosis reagent

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Effect test

Embodiment 1

[0034] Example 1 Screening and identification of lncRNA-NEAT1

[0035] The inventors of the present invention screened 202 common lncRNAs from the accumulated 1500 cases of glioma databases CGGA, Rembrandt, TCGA and GSE16011, and found that 6 lncRNAs were highly expressed in glioma EGFR module type, and in the above-mentioned The 6 lncRNAs were verified by PCR in the U87-MG cell line in the 4 major glioma authoritative public databases, and it was found that the expression of lncRNA-NEAT1 was the highest in the U87-MG cell line (see figure 1 ).

[0036] In situ hybridization was used to detect the expression level of lncRNA-NEAT1 in glioma tissues of patients with PDGFR module and EGFR module, and the results were as follows figure 2 shown.

[0037] Then, the receiver operating characteristic curve (ROC) of NEAT1 in the EGFR module was evaluated in the above four authoritative public databases. The results are as follows: image 3 shown.

[0038] These results show that l...

Embodiment 2

[0039] Example 2 Application of lncRNA-NEAT1 as a molecular pathological diagnostic marker in the detection of EGFR module glioma

[0040] 1. Collection, processing and extraction of RNA from tumor tissue samples

[0041] (1) Collection of glioma tissue samples: pathological tissues of glioma patients were collected.

[0042] (2) Treatment of glioma tissue samples: Rinse with PBS and store in liquid nitrogen.

[0043] (3) Extraction of RNA in glioma tissue: 100 mg of each tissue sample was taken, ground into powder, and RNA was extracted according to Trizol purchased from Sigma Company.

[0044] (4) Reverse transcription into cDNA: take the same amount of RNA from each sample, and perform reverse transcription with random primers. According to 5×M-MLVRT Buffer 5μl, MgCl 2 2 μl, M-MLV Reverse transcript 1 μl, rRNasin 0.5 μl (Promega), dNTPs 1 μl, random primer 1 μl, extracted RNA 2ul, DEPC water (supplemented with H 2 O to bring the total volume to 20 μl) for reverse trans...

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Abstract

The invention discloses application of lncRNA-NEAT1 as a molecular pathological diagnosis marker in preparation of a glioma diagnosis reagent. According to the invention, a glioma public database is utilized for screening lncRNA related to EGFR module and PDGFR module type gliomas, and detection on tumor tissues of a glioma patient finds that lncRNA-NEAT1 can be detected in the tumor tissues, andthe lncRNA-NEAT1 content of the EGFR module tumor tissues is obviously higher than that of PDGFR module, so that the marker can be used for distinguishing the EGFR module type glioma and the PDGFR module type glioma. Besides, the invention also provides a glioma diagnosis kit and establishes a glioma diagnosis method. The method is carried out by virtue of a PCR process, has the advantages of lowcost, short time and high sensitivity compared with the traditional method and can meet large-scale experiment and clinical application needs.

Description

technical field [0001] The present invention relates to a molecular pathological diagnostic marker for detecting glioma, in particular to the use of lncRNA-NEAT1 as a molecular pathological diagnostic marker in the preparation of glioma diagnostic reagents, and also relates to a glioma diagnostic kit. The invention belongs to the technical field of medicine. Background technique [0002] The classification of tumors provides an important direction for the clinical treatment and scientific research of tumors. At present, the classification and diagnosis of gliomas (the World Health Organization standard in 2007) widely used in the medical field are largely based on morphological indicators. This classification system not only fails to reveal the biological nature of tumor development, but also is highly subjective, with inconsistencies as high as 40%. [0003] Molecular pathology is the use of molecular and genetic approaches to diagnose and classify tumors, design and valid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6851C12Q1/6886C12Q2600/158C12Q2600/178C12Q2531/113C12Q2563/107
Inventor 蔡金全陈群康春生蒋传路
Owner HARBIN MEDICAL UNIVERSITY
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