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Application of protein Bach1 to preparation of cell Wnt signaling pathway inhibitor

A technology of signaling pathway and inhibitor, applied in the fields of biotechnology and medicine

Active Publication Date: 2017-01-04
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is known that the downstream genes regulated by Bach1 include heme oxygenase-1 (heme oxygenase-1, HO-1), etc. However, there are no reports about Bach1 as an inhibitor of Wnt signaling pathway and related applications.

Method used

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  • Application of protein Bach1 to preparation of cell Wnt signaling pathway inhibitor
  • Application of protein Bach1 to preparation of cell Wnt signaling pathway inhibitor
  • Application of protein Bach1 to preparation of cell Wnt signaling pathway inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Bach1 inhibits cell Wnt / β-catenin regulatory pathway test

Embodiment 2

[0058] Example 2: Bach1 induces cell cycle arrest and apoptosis, inhibits cell proliferation assay

[0059] (1) Cell cycle analysis experiment

[0060] Use Bach1 adenovirus or control adenovirus to infect human microvascular endothelial cells, and Bach1 adenovirus (Ad-Bach1) or control virus (Ad-GFP) is provided by Shanghai Jikai Gene Chemical Technology Company; after 72 hours, collect cells, wash with ice ethanol After fixing, discard the supernatant after centrifugation, add 1ml DNA staining solution (containing PI and RNaseA), incubate at room temperature for 30 minutes, and send to the flow cytometer for detection;

[0061] (2) Apoptosis experiment

[0062] Use the Annexin V / PI kit (Invitrogen, USA) to detect cell apoptosis, collect human microvascular endothelial cells, add 5 μl Alexa per 100 μl cell suspension 488 annexin V (Component A) and 1 μl 100 μg / ml PI working solution; incubate at room temperature for 15 minutes; add 400 μl 1×annexin-binding buffer (Component...

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Abstract

The invention belongs to the fields of biotechnology and medicine, and relates to application of protein Bach1 to inhibition of a cell classic Wnt signaling pathway. The invention provides that the Bach1 is a medicine target for intervening diseases related to an abnormal Wnt signaling pathway, such as vasculogenesis, cancers, abnormal cell proliferation, Alzheimer's disease and osteoarthritis. The protein Bach1 directly blocks beta-catenin / TFC protein binding to interfere formation of the transcription complex so that Wnt signaling pathway target gene expression is inhibited; the cell proliferation is inhibited and apoptosis is induced. The application provided by the invention provides a new target for screening a medicine for treating the diseases related to the abnormal Wnt signaling pathway; furthermore, the protein Bach1 can be used for preparing a cell Wnt signaling pathway inhibitor and preparing a medicine for preventing and / or treating the diseases related to the abnormal Wnt signaling pathway.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and relates to a new application of protein Bach1, in particular to an application of protein Bach1 in preparing inhibitors of cell Wnt signaling pathways. Background technique [0002] The prior art discloses that the Wnt signaling pathway is a very important signaling pathway in cells, which is closely related to tumors, angiogenesis, cell proliferation, tissue regeneration, embryonic development, and the occurrence and development of nerves and bones. Studies have shown that the Wnt signal transduction pathway is mainly divided into the canonical Wnt signal transduction pathway (Wnt / β-catenin pathway) dependent on the β-catenin / TCF transcription complex and the non-dependent Wnt signal transduction pathway independent of the β-catenin / TCF transcription complex. Canonical Wnt signaling pathway (Wnt / Ca 2+ pathway and Wnt / JNK pathway). In the canonical Wnt signal transduction pathway,...

Claims

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Application Information

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IPC IPC(8): C07K14/47A61K38/17A61P35/00A61P9/00A61P9/10A61P11/00A61P29/00A61P19/02A61P31/10A61P31/12A61P19/08A61P25/28A61P17/00
CPCA61K38/1709C07K14/4703
Inventor 孟丹蒋丽牛琮魏香香陈思锋王新红刘俊许赵凤娣
Owner FUDAN UNIV
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