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Application of polypeptide in preparation of drug for preventing and treating glioblastoma

A brain glioma and drug technology, which is applied in the potential medical application of polypeptide and its preparation field, and achieves the effect of good application prospect

Active Publication Date: 2015-06-24
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no relevant report about alleviating the pathological process of glioma by inhibiting the activity of TROY receptor by polypeptide

Method used

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  • Application of polypeptide in preparation of drug for preventing and treating glioblastoma
  • Application of polypeptide in preparation of drug for preventing and treating glioblastoma
  • Application of polypeptide in preparation of drug for preventing and treating glioblastoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Polypeptide inhibits the combination of TROY and downstream signaling molecule RKIP and reduces the activity of NF-kB

[0032] 1. Experimental materials

[0033] 1. Strains

[0034] Escherichia coli DH5α, BL21(DE3) strains were provided by our laboratory.

[0035] 2. Plasmid

[0036] The pTAT vector was obtained from the laboratory of Professor Dowdy SF, and the pGEX-4T3-1 vector was provided by our laboratory.

[0037] 3. Tool enzyme

[0038] Restriction endonuclease, T4DNA ligase, TaqDNA polymerase and other tool enzymes were purchased from Shanghai Huamei Bioengineering Company, Shenggong Company and Sigma Company.

[0039] 4. Animals

[0040] 10-20g nude mice were purchased from Shanghai Slack Experimental Animal Center.

[0041] 5. Kit

[0042] The column centrifugal gel extraction kit was purchased from Shanghai Huashun Biological Reagent Company.

[0043] 6. Medium

[0044] LB medium and NZCYM medium preparation reagents were purchased from DI...

Embodiment 2

[0161] Embodiment 2: Polypeptide is used for the animal experiment of treating glioma

[0162] Tumor formation experiment in nude mice Planting experiment is a commonly used animal model to study the process of tumor lesions. We first cultured the U87 glioma cell line, digested and centrifuged the cells and suspended the cells in serum-free DMEM to make 1×10 7 / ml of U87 cell suspension with a density of about

[0163] The TAT-TROY interfering peptide purified in the above experimental method 13 was injected into the peritoneal cavity of the mouse, the growth of the tumor mass was observed according to the time course, and the length and width data were measured for statistical analysis. Through the above studies, the role of TROY / RKIP interaction in glioma was further clarified.

[0164] 1. Experimental materials

[0165] Nude mice, male, weighing 10-20 g, 5 mice / cage. Purchased from Shanghai Xipuer-Pika Laboratory Animal Co., Ltd. (production license number: SCXK (Shangh...

Embodiment 3

[0170] Embodiment 3: The experiment of shRNA

[0171] 1. Experimental method

[0172] 1) Inoculate 1×10 5 Put each U87 cell into a 6-well culture plate, add a medium volume of 5 ml to each well, and the confluence of the cells during virus infection is 50%.

[0173] 2) Change the medium for the cells before infection, suck off the supernatant of the cells, and add the required culture medium according to different groups.

[0174] 3) Add the control and TROY-targeted lentivirus solutions to the culture wells, and the shRNA sequences designed for the human TROY (NM_018647) sequence are:

[0175] 1#, 5-ATCAACTCAGGATGCACTA-3 (SEQ ID NO: 3),

[0176] 2#, 5-TCAACGTCTTTGGATTCAA-3 (SEQ ID NO: 4),

[0177] 3#5-AGGCTATTTGTCATGTAAA-3 (SEQ ID NO:5),

[0178] The control was 5-TTCTCCGAACGTGTCAC-GT-3 (SEQ ID NO: 6).

[0179] 4) Put the cells back into the incubator and incubate for 8-12 hours to observe the state of the cells. Continue culturing and replace with fresh medium after 2...

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Abstract

The invention relates to the technical field of biological medicines, in particular to an application of a polypeptide in preparation of a drug for preventing and treating glioblastoma. The polypeptide has the functions of inhibiting cell proliferation of glioblastoma and delaying a growing process of glioblastoma, is peptide fragments (SEQ ID NO:1) (Sequence Identifier Number:1) of 138 amino acids and positioned at NO 234 to 371 amino acids in a cell interior domain of a human TROY protein sequence, has the functions of blocking the combination of a TROY accepter with a downstream signaling molecule RKIP (Phosphatidylethanolamine Binding Protein), reducing TROY mediated NF-kB activation, and inhibiting the cell multiplication of glioblastoma.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the potential medical application and preparation of a polypeptide capable of inhibiting the proliferation of brain glioma cells and delaying the growth process of brain glioma. Background technique [0002] At present, there is no effective treatment for brain tumors: surgical resection is not only difficult, has many side effects, but also is prone to recurrence; the existence of the blood-brain barrier makes it difficult for most chemotherapy drugs to reach the brain lesion and play an effective role; most brain tumors are not sensitive to radiotherapy. Therefore, exploring new methods for the diagnosis and treatment of brain tumors and effectively improving the quality of life of patients has become one of the medical problems that basic researchers and clinicians in the field of neuroscience need to solve urgently. [0003] Glioma accounts for about 30%-80% of all brain ...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61K48/00A61P35/00
Inventor 何成刘秀杰
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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