Wnt-targeted drug screening cell model, its construction and application

Abstract

The invention provides a Wnt-targeted drug screening cell model, which is constructed by using a Wnt specific binding sequence-containing report system vector and taking a constitutive Wnt-containing active cell as the basis. The model is used for screening drugs treating Wnt constitutive activation caused tumors and immunological diseases. Due to employment of cells with constitutive Wnt activity, the reporter gene in model cells is in a highly activated state with no need to use additional irritant, and the obtained stable cell model can be directly used for compound screening. According to the invention, the screening cost is greatly reduced, and also, the whole selection process is simplified from cell culture, Wnt activation, compound adding, and detection to cell culture, compound adding, and detection. With the model, the screening cycle is shortened, the operation steps are reduced, and the system stability, high efficiency and usability are enhanced. Thus, the cell model is more suitable for high-throughput drug screening.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to a drug screening cell model with Wnt as the target, in particular to a drug screening cell model based on cells with constitutive Wnt activity; the invention also relates to the drug screening cell model Build methods and applications. technical background [0002] Wnt is an evolutionarily highly conserved signaling pathway, and members of the Wnt family exist from nematodes to humans. Drosophila contains seven Wnt genes, compared with 19 in vertebrates. The experiment of β-catenin (CTNNB1) function deficiency and gain of function in mice confirmed that at various stages of early embryonic development, members of the Wnt family directly determine the direction of cell differentiation, and also regulate the regulation of cardiovascular, central nervous system, etc. Differentiation and development of tissues and organs such as kidney and lung. In adults, Wnt signaling maintains tissue sel...

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Problems solved by technology

[0004]Based on the above reasons, researchers have been trying to find Wnt inhibitors with good specificity and low side effects, so that they can be finally used clinically to alleviate and treat a large number of related diseases. patients; however, there are very few ideal products today, largely due to the lack of an efficient and simple screening method; although the high-throughput screening method based on the reporting system has appeared and been used in practice, it has achieved certain results. However, there are still some insurmountable shortcomings, such as the need for exogenous stimulation (activating Wnt with an activator first, and then screening for inhibitors), which not only requires many steps, but also has high screening costs, especially for cytokines such as Wnt3 The use of activators will greatly increase the cost; even if the cost of activators like Licl is low, but due to its high toxicity, it limits Its use; at the same time, each additional step in the screening system will increase the instability of the reporting system (additional operations will inevitably bring additional uncertainties)

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