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Screening method of drugs influencing classic Wnt signal pathway, and its application

A screening method and drug technology, applied in the direction of testing pharmaceutical preparations, material inspection products, etc., can solve the problem of no obvious progress in research.

Active Publication Date: 2014-09-10
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Since the Wnt / β-catenin signaling pathway is related to many cancers and diseases, the out-of-control Wnt / β-catenin signaling pathway can be used as a good way to treat diseases related to the Wnt / β-catenin signaling pathway, but at this stage There has been no significant progress in the research

Method used

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  • Screening method of drugs influencing classic Wnt signal pathway, and its application
  • Screening method of drugs influencing classic Wnt signal pathway, and its application
  • Screening method of drugs influencing classic Wnt signal pathway, and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] 1. Experimental materials

[0077] 1) Plasmid

[0078] Axin-HA and GSK3β-Myc were obtained by PCR and cloned into mammalian expression vectors pCMV-HA and pCMV-Myc, respectively. (Refer to Chen, T., et al., Identification of zinc-finger BEDdomain-containing3 (Zbed3) as a novel Axin-interacting protein that activates Wnt / beta-catenin signaling. The Journal of biological chemistry, 2009.284 (11): p .6683-9 operation)

[0079] LRP6 and MESD plasmids (expression vector is pCS2+) were donated by Prof. Xi He (Children's HospitalBoston, USA). The two plasmids can be referred to Li, Y., et al., Modulation of LRP6-mediated Wnt signaling by molecular chaperone Mesd. FEBS letters , 2006.580(22): p.5423-8 prepared.

[0080] TOPFlash plasmid was purchased from Millipore Corporation.

[0081] GFP plasmid was purchased from Millipore Corporation.

[0082] 2) Antibody

[0083] The antibody of HA (1666606, Roche) was purchased from Roche; the antibody of LRP6 (#3395, Cell signalin...

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Abstract

The invention provides a screening method of drugs influencing a classic Wnt signal pathway, and its application. The screening method of drugs influencing a classic Wnt signal pathway treats Axin and LRP5 / 6 interaction as a drug screening target. The drug screening method can be used for screening drugs for preventing or treating diseases or imbalance caused by the abnormity of the classic Wnt signal pathway, drugs for preventing or treating diseases or imbalance needing specific activation or inhibition of the classic Wnt signal pathway, or stem cell number amplification drugs.

Description

technical field [0001] The present invention relates to a drug screening method, in particular to a drug screening method affecting canonical Wnt signaling pathway and application thereof. Background technique [0002] The Wnt signaling pathway plays an important regulatory role in the early embryonic development of organisms, and can participate in the regulation of body axis formation, organogenesis and cell fate determination and other physiological processes (Clevers, H. and R. Nusse, Wnt / beta- Catenin Signaling and Disease. Cell, 2012.149(6):p.1192-205). Once the Wnt signaling pathway is blocked, animal embryos will produce very obvious mutant phenotypes due to the disruption of the morphological process, such as abnormal epidermis in Drosophila, ventralized limbs in mice, loss of asymmetric division of EMS cells in nematodes, and so on. In addition, the occurrence of many tumors is related to the mutation of Wnt signaling pathway members, such as the disorder of APC p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/15
Inventor 李林汪胜郝小江宋晓敏尹俊林陈铎之
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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