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Screening system of anti-tumor drugs based on TRAIL receptor signal channel

An anti-tumor drug and signaling pathway technology, applied in the field of anti-tumor drug screening system based on TRAIL receptor signaling pathway, can solve the problem of no anti-tumor drug screening system, and achieve high novelty and practicability, high reliability and Adaptability, cost and time saving effects

Inactive Publication Date: 2014-03-12
SHANDONG UNIV
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After searching, there is currently no report on an anti-tumor drug screening system based on the TRAIL receptor signaling pathway

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  • Screening system of anti-tumor drugs based on TRAIL receptor signal channel
  • Screening system of anti-tumor drugs based on TRAIL receptor signal channel
  • Screening system of anti-tumor drugs based on TRAIL receptor signal channel

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Embodiment 1

[0038]The antitumor drug screening system based on the TRAIL receptor signaling pathway of the present invention is composed of cells containing a fusion gene of DR5 promoter and luciferase reporter gene, a cell culture lysis reagent, and a luciferase detection substrate. It is mainly constructed by molecular cloning technology and lentiviral expression system.

[0039] The construction and application steps of the anti-tumor drug screening system based on the TRAIL receptor signaling pathway are as follows:

[0040] 1) Cloning of DR5 promoter gene

[0041] Using the cDNA of A549 cells as a template, the DR5 promoter gene was cloned by PCR technology according to the Invitrogen PCR kit instructions (such as figure 1 ). Then, under the action of T4 DNA ligase, it was ligated with the pGM-T vector at 16°C for 16 hours, and prepared Escherichia coli DH5α competent cells according to the molecular cloning experiment guideline. Bath for 30 minutes, heat shock at 42°C for 90s, ad...

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Abstract

The invention discloses a screening system of anti-tumor drugs based on a TRAIL receptor signal channel. The screening system comprises fusion gene cells (A549-pDR5-luc and H157-pDR5-luc), CCLR and luciferase detection substrate, wherein the fusion gene cells comprise a DR5 promoter and a luciferase reporter gene. According to the invention, through determining the rise of luciferase reporter gene expression in the A549-pDR5-luc and H157-pDR5-luc cells on which a compound to be screened acts to screen a compound improving the expression of DR5, the screening of anti-tumor drugs is realized. The screening system has characteristics of short experimental period, quick detection process, high sensitivity and wide application, and provides a new method for the screening of high through-put anti-tumor drugs.

Description

technical field [0001] The invention relates to an anti-tumor drug screening system, in particular to an anti-tumor drug screening system based on TRAIL receptor signaling pathway. Background technique [0002] Cancer has become one of the most lethal diseases of human beings, seriously threatening people's life expectancy. The mortality rate of malignant tumors among urban and rural residents in my country is already at a high level in the world, and it shows a continuous growth trend. There are 2.2 million new cancer cases and 1.6 million cancer deaths in China every year. Among them, lung cancer and breast cancer have increased by 465% and 96% respectively in the past 30 years. Malignant tumor has become the first cause of death of urban residents in my country, and the second cause of death of rural residents. Therefore, our research on tumor prevention and treatment is urgent. [0003] Chemotherapy is one of the important methods to treat cancer at present, and it i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/66C12Q1/02
Inventor 苏玲刘相国徐林艳杨丽娜
Owner SHANDONG UNIV
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