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Reagent for blocking interaction between Netrin-1 and CD146 and tumor treatment application thereof

A technology of uses and reagents, applied in the field of reagents for blocking the interaction between Netrin-1 and CD146 and their use in tumor therapy, which can solve the problems of unclear mechanisms, unknown receptors and downstream signals, etc.

Inactive Publication Date: 2014-10-08
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism by which Netrin-1 promotes angiogenesis is still unclear, and its receptors and downstream signals are unknown. Therefore, there is currently no anti-tumor angiogenesis drug targeting Netrin-1.

Method used

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  • Reagent for blocking interaction between Netrin-1 and CD146 and tumor treatment application thereof
  • Reagent for blocking interaction between Netrin-1 and CD146 and tumor treatment application thereof
  • Reagent for blocking interaction between Netrin-1 and CD146 and tumor treatment application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Interaction of Netrin-1 and CD146 at cellular and in vitro levels

[0033] CD146, as an endothelial cell adhesion molecule, plays a key role in angiogenesis, especially pathological angiogenesis. Netrin-1 has also been reported to promote angiogenic processes, but its receptors are not well understood. In order to study the relationship between the two, we confirmed the direct interaction between Netrin-1 and CD146 at the level of cells and in vitro by co-immunoprecipitation and other methods. Monoclonal antibodies against CD146 can block the interaction between Netrin-1 and CD146.

[0034] Main material: human embryonic kidney epithelial cells (HEK293) (ATCC, CRL-1573).

[0035]Main reagents: cell lysate, PBS, HEPES. Mouse-derived anti-CD146 monoclonal antibody AA1 (produced independently by our laboratory. Preservation institution: China General Microbiology Collection Center; preservation number: CGMCC No.2310; preservation date: 2007.12.28). Mouse ant...

Embodiment 2

[0062] Example 2: Netrin-1-induced endothelial cell proliferation, migration and angiogenesis depend on the binding of Netrin-1 to the receptor CD146

[0063] Angiogenesis mainly provides nutrients for tumors and provides pathways for tumor cell metastasis, so it plays a vital role in tumor growth. Inhibition of angiogenesis can inhibit tumor growth to a great extent. Netrin-1 promotes the proliferation, migration and angiogenesis of endothelial cells by binding to the receptor CD146 on the surface of endothelial cells, and anti-CD146 monoclonal antibodies inhibit Netrin-1-induced endothelial cells by blocking the binding of Netrin-1 to CD146 Function.

[0064] Main materials: human umbilical vein endothelial cell line (HUVEC, purchased from CellSystems Biotechnolegie Vertrieb), 96-well transwell plate (Corning HTS Transwell-96Cell Migration Products).

[0065] Main reagents: recombinant human Netrin-1 protein, CD146-specific siRNA (synthesized in Invitrogen Company), anti-C...

Embodiment 3

[0084] Example 3: CD146-specific siRNA or anti-CD146 monoclonal antibody AA98 can block the activation of signaling pathways caused by Netrin-1 stimulation

[0085] In endothelial cells, Netrin-1 binds to the receptor CD146 and can activate downstream signaling molecules such as ERM, p38, ERK, and NF-κB. CD146-specific siRNA can knock down the expression of CD146 on endothelial cells and inhibit the downstream signaling pathway induced by Netrin-1. Similarly, antibody AA98 can inhibit the downstream signaling of Netrin-1 activation by blocking the binding of Netrin-1 to CD146.

[0086] Main materials: human umbilical vein endothelial cell line.

[0087] Main reagents: recombinant human Netrin-1 protein, CD146-specific siRNA, anti-CD146 antibody AA98, mouse IgG, etc.

[0088] Main method: HUVEC cells transfected with CD146-specific siRNA or control siRNA were stimulated with Netrin-1 (50ng / ml) and then lysed for biochemical analysis of signaling pathways (A). Or use anti-CD1...

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Abstract

The invention discloses an application of a reagent for blocking interaction between Netrin-1 and CD146 in preparing medicaments for inhibiting neovascularization and an application of the reagent in preparing medicaments for treating tumors, as well as a method for inhibiting neovascularization and treating tumors by blocking interaction between Netrin-1 and CD146.

Description

technical field [0001] The present invention relates to the use of a reagent for blocking the interaction between Netrin-1 and CD146 in the preparation of medicines for inhibiting neovascularization and the use in the preparation of medicines for treating tumors. The present invention also relates to a method for inhibiting neovascularization and treating tumors by blocking the interaction between Netrin-1 and CD146. Background technique [0002] Cancer is currently the number one killer that endangers human life and health. The annual incidence of malignant tumors in my country is about 3.12 million. Six people are diagnosed with cancer every minute in the country, and those who die from cancer account for more than 20% of the total number of deaths, ranking first in the cause of death of urban residents (according to 2012 statistics). [0003] Currently, drug therapy is an important aspect in the three conventional (radiotherapy, chemotherapy, and surgery) treatments for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K39/395A61P35/00
CPCA61K39/3955
Inventor 阎锡蕴凃滔杨东玲冯静宋丽娜晏慧文
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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