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Oligoden droglioma (OG) marker MAP2 (microtubuleassociated protein 2) protein and use thereof

A glioma, marker technology, applied in the field of biotechnology and medicine, can solve the problems of cumbersome process and high cost

Inactive Publication Date: 2012-04-11
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection method of 1p allele deletion commonly used at present has: FISH (fluorescence in situ hybridization) and microsatellite DNA method, and described method needs special equipment and technology, and often process is loaded down with trivial details, and expense is expensive, and wherein, microsatellite DNA method is more It requires the patient's genomic DNA (obtained from the patient's blood) other than the patient's tumor tissue sample, which introduces more cumbersome steps such as collection and processing

Method used

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  • Oligoden droglioma (OG) marker MAP2 (microtubuleassociated protein 2) protein and use thereof
  • Oligoden droglioma (OG) marker MAP2 (microtubuleassociated protein 2) protein and use thereof
  • Oligoden droglioma (OG) marker MAP2 (microtubuleassociated protein 2) protein and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Preparation of tissues and specimens (oligodendroglioma frozen specimens and paraffin-embedded specimens)

[0044] The isolated fresh tumor specimens obtained from the surgery were quickly frozen and stored at -80°C (all oligodendroglioma specimens and clinical data of patients were obtained from Fudan University after obtaining the informed consent of the patients in accordance with the protocol approved by the Institutional Review Board. University Affiliated Huashan Hospital). Histological evaluation ensures that the obtained tumor tissue contains more than 80% tumor cells to facilitate proteomic studies. An additional 39 formalin-fixed and paraffin-embedded tumor specimens were used for the identification of candidate proteins. Tumor grades were performed by two neuropathologists according to "WHO Histological Classification (2000)".

Embodiment 2

[0045] Example 2: Analysis of three-color FISH technique to detect the deletion of alleles on chromosome arm 1p

[0046] Cut 5 μm thick frozen sections, fix with 100% alcohol, and fix with 0.2% paraformaldehyde / PBS for 10 minutes at 4°C. Use commercialized 1p36 microdeletion syndrome-specific three-color fluorescently labeled probes, namely LSI 1p36, TelVysion 1p and LSI 1q25 probes labeled with orange, green and light blue-green three-color fluorescent isotopes, respectively, and perform fluorogens according to the operation manual. Site hybridization to detect the deletion of alleles on chromosome arm 1p: it includes brief steps: denature the section at 73°C for 5 minutes, add 10 μl of probe mixture, cover with a coverslip, incubate overnight at 37°C, wash at 45°C, and transfer to 2 ×SSC / 0.1% NP-40 solution (room temperature). Then stained by DAPI-II. When paraffin-embedded sections are used for three-color FISH analysis, they need to be boiled in sodium citrate buffer for...

Embodiment 3

[0049] Example 3: Proteomic analysis of protein profiles based on iTRAQ-LC-MS / MS technology, looking for differential proteins

[0050] In the iTRAQ experiment of the present invention, high-abundance proteins are directly obtained from isolated tumor tissues, and the obtained differentially expressed proteins directly represent the tumor itself and the tumor microenvironment related to chemotherapy tumor biology.

[0051] iTRAQ sample preparation: Select 8 frozen samples of oligodendroglioma (4 of which are 1p heterozygous deletion, 4 without 1p heterozygous deletion) for proteome sequence analysis, take 50mg of solid tumor tissue, and lysate (Tris-HCl 50mM, thiourea 2.5M, urea8M, CHAPS 4%, DTT 65mM) and mix well to extract the total protein. Sonication, 6×10s pulse, operation on ice. Centrifuge at 20000g for 45min to remove cell debris. The entire operation of this experiment was performed on ice. Use the Ready Prep 2-D Cleanup Kit (Bio-Rad Laboratories, Inc.USA) to purif...

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Abstract

The invention belongs to the field of biotechnology and medicine, relates to oligoden droglioma (OG) markers, and particularly relates to a protein marker MAP2 (microtubuleassociated protein 2) protein which can differentiate OG that clinically has sensitivity to chemotherapy and 1P loss of heterozygosity (1P LOH), and use thereof. According to the invention, the protein marker MAP2 protein which can differentiate the OG that clinically has sensitivity to chemotherapy and 1P loss is determined through the iTRAQ-LC-MS / MS technology. The MAP2 protein has 90% accuracy for the prognosis of the 1P loss. The marker not only can explain the mechanism for the sensitivity to chemotherapy of an OG sample with the 1P LOH, but also can prognose the sensitivity to chemotherapy of an OG patient with the 1P LOH through a decision tree model. The MAP2 protein can be further used for preparing a clinical kit so as to offer help for the diagnosis, the prognosis and the treatment of the OG.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine. It relates to markers of human oligodendroglioma, in particular to a human oligodendroglioma (Oligoden droglioma) capable of distinguishing clinically chemotherapy-sensitive, 1p allele heterozygous loss (1p Loss of heterozygosity, 1p LOH) , OG) protein marker MAP2 protein (microtubulea associated protein 2, MAP2) and its use. Background technique [0002] Biomarkers refer to physical, chemical, or biological indicators that can be objectively measured, indicating normal physiological or pathological processes, or the body's response to drugs. One of the most critical issues in modern tumor research is to find markers for early detection, diagnostic typing, treatment monitoring and prognosis, and drug targets. Studies have revealed a series of changes in the intracellular protein expression profile during the transition from a normal state to a tumor. Tumor markers are a class of subs...

Claims

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Application Information

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IPC IPC(8): C07K14/47G01N33/53G01N27/62G01N21/64G01N27/626
Inventor 施前杨柳松徐晓恩
Owner FUDAN UNIV
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