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Method for determining APRIL (A Proliferation-inducing Ligand) gene promoter and transcription factor binding site and use

A technology of transcription factors and binding sites, which can be used in biochemical equipment and methods, medical preparations containing active ingredients, drug combinations, etc., and can solve problems such as unclear

Inactive Publication Date: 2012-07-04
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is still unclear which receptor has a direct effect on the APRIL signaling pathway

Method used

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  • Method for determining APRIL (A Proliferation-inducing Ligand) gene promoter and transcription factor binding site and use
  • Method for determining APRIL (A Proliferation-inducing Ligand) gene promoter and transcription factor binding site and use
  • Method for determining APRIL (A Proliferation-inducing Ligand) gene promoter and transcription factor binding site and use

Examples

Experimental program
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Embodiment Construction

[0035] Cell culture:

[0036] The colon cancer SW480 cell line was cultured in a single layer in a complete medium containing 10% fetal bovine serum, digested and passaged in 0.25% trypsin at 37°C in a constant temperature incubator with 5% CO2 saturated humidity. The number of cells is 2×10 6 Cells / ml subcultured into a 6-well culture plate. When the cell confluence reached over 90%, transfect the plasmid according to the instructions of Liposome 2000, and observe the transfection effect under an inverted fluorescent microscope.

[0037] Promoter region and transcription factor analysis:

[0038] Taking the 3.5kb of the 5'flanking region of the APRIL gene as the research object, the potential promoter prediction software Promoter 2.0Prediction Server (http: / / www.cbs.dtu.dk / services / Promoter / ) and Promoter Scan program (http: / / www-bimas.cit.nih.gov / molbio / proscan / ) for analysis. CpG islands were searched using CpG Island Searcher (http: / / cpgislands.usc.edu / ). The potential ...

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Abstract

The invention discloses a method for determining an APRIL (A Proliferation-inducing Ligand) gene promoter and transcription factor binding site and use. According to the invention, a promoter region of APRIL is cloned and identified. As discovered, a region from -1539 to -1001 has an important effect on transcription adjustment and control over APRIL; a transcription factor Sp1 or NF-kB exits in the region; and the transcription of the APRIL gene can be activated. The invention provides application of mithramycin and Bay11-7082 to preparation of medicaments for treating human colorectal adenocarcinoma; and the effect is clear.

Description

technical field [0001] The invention relates to a method and application for determining human APRIL gene promoter and transcription factor binding site. Background technique [0002] Worldwide, colorectal cancer ranks third among cancer-related deaths. In recent years, the incidence of colorectal cancer in Asian and Eastern European countries has been on the rise. APRIL was cloned and identified in 1998. In addition to the high expression of APRIL in colorectal cancer, lymphoma and melanoma cells, the expression level of APRIL is very low in other tissues, indicating that APRIL has tissue specificity. Previous studies found that APRIL was highly expressed in the colorectal cancer cell line SW480 compared with its surrounding normal tissues. Studies in our laboratory also found that APRIL was highly expressed in colorectal cancer SW480 cells. In addition, the APRIL interference plasmid was transfected into the SW480 colorectal cancer nude mouse model, and the tumors in t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/66A61K31/704A61K31/277A61P35/00
Inventor 王惠民徐锦景蓉蓉季伙燕
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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