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Cell model for quick screening of histone deacetylase inhibitor

A deacetylase and inhibitor technology, which is applied in the field of cell models for rapid screening of histone deacetylase inhibitors, and can solve problems such as unfavorable gene transcription and gene expression shutdown.

Inactive Publication Date: 2007-08-01
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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Problems solved by technology

The deacetylation of histones makes the DNA and histones tightly bound, which is not conducive to gene transcription and leads to the shutdown of some genes.

Method used

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  • Cell model for quick screening of histone deacetylase inhibitor

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Embodiment Construction

[0020] The following combination of different types of HDAC inhibitors Depsipeptide (FK228), suberoylanilidehydroxamic acid (SAHA), Trichostatin A (TSA) and other 4 kinds of drugs with anti-tumor activity 5-aza-2'-deoxycytidine (DAC) , retinoic acid, paclitaxel and 17-AAG respectively treated the experiment of the cell model to further illustrate the present invention.

[0021] 1. Experimental method

[0022] 1.1 Using the method of molecular cloning, the above-mentioned promoter sequence fragments were respectively inserted into the commercial vectors containing the luciferase reporter gene. After plasmid amplification and purification, the COS7 cells were transfected with liposome-mediated method, G418 (1mg / ml ) After pressurized selection for 21 days, a cell-resistant clone stably expressing the foreign gene was obtained, and the resistant cell could be used as a model for subsequent drug screening.

[0023] 1.2 HDAC inhibitors FK228 and TSA were provided by Dr. David Schr...

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Abstract

The invention discloses a cell sifting moudle for fast sifting of antineoplastic- histone precursor histone deacetylase inhibitor compound from compound storage during new drug development. The moudle makes use of speciality that the precursor histone deacetylase inhibitor can activate some starting subsequence, constructs te oexpression vector containing special starting factor and firefly luciferase reporting gene, and builds cos- 7 clone that stably expresses said vector. The cell contains starting subsequence with different degree of activity for luciferase, the starting factor can be acitvated by histone precursor histone deacetylase inhibitor and then the expression of luciferase reporting gene is intensified, and so the detection for histone precursor histone deacetylase inhibitor activity in sample through detection of luciferase activity change can be finished in a short time. The cell sifting moudle can be used for fast and high- flux sifting of new antineoplastic and histone precursor histone deacetylase inhibitor compound from compound storage during new drug development.

Description

technical field [0001] The invention belongs to a cell model used in the research and development of new compound drugs that can perform rapid and high-throughput screening on compound libraries to obtain lead compounds of histone deacetylase inhibitors. Background technique [0002] Histones are an important part of the basic components of human chromatin, consisting of DNA double-strand winding histone heterooctamers. Studies have shown that there are various modifications at the lysine tail of histones, including phosphorylation, methylation, acetylation, and ubiquitination, etc. These modifications play an important role in regulating gene transcription and expression. It is known that there are two enzymes involved in the regulation of histone acetylation in cells, that is, histone acetyltransferase, which can enhance acetylation, and histone deacetylase (HDAC), which can inhibit acetylation. Under normal conditions, a dynamic balance between the activities of these tw...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68C12N15/113
Inventor 于晓妉杜芝燕王妍徐元基
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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