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Mononucleotide polymorphism of human heterogenous substance metabolism enzyme gene and application of diagnosing and treating large intestinal cancer

A single nucleotide polymorphism, polymorphism technology, applied in the screening of appropriate therapies for different types of colorectal cancer, diagnosis and treatment of this common cancer field, can solve problems such as narrowing of enzyme activity and difficulty in safe use of doses

Inactive Publication Date: 2007-03-14
PHARMACOGENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The narrowing of CYP2C9 enzyme activity makes it very difficult to estimate the safe dosage of some drugs, especially some drugs with narrow therapeutic index, such as S-warfarin, tolbutamide and phenytoin (Schwarz UI., 2003.Clinical relevance of genetic polymorphisms in the human CYP2C9 gene. Eur J Clin Invest. 33 Suppl 2:23-30)

Method used

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  • Mononucleotide polymorphism of human heterogenous substance metabolism enzyme gene and application of diagnosing and treating large intestinal cancer
  • Mononucleotide polymorphism of human heterogenous substance metabolism enzyme gene and application of diagnosing and treating large intestinal cancer
  • Mononucleotide polymorphism of human heterogenous substance metabolism enzyme gene and application of diagnosing and treating large intestinal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Example 1: Detection of the newly discovered single nucleotide polymorphism site 51523T>G in the present invention

[0111] The present invention has newly discovered a single nucleotide polymorphism site 51523T>G located on intron 9 of the ABCG2 gene, and the method for detecting this site is introduced as follows:

[0112] Available software Primer3 ( http: / / frodowi.mit.edu / cgi-bin / primer3 / primer3 ) Design some specific primers to search for polymorphic sites in related regions. use BLAST TM The program (National Center for Biotechnology Information - http: / / www.ncbi.nlm.nih.gov / BLAST) tested the specificity of the designed primers relative to the human genome sequence. Regardless of the forward or reverse primers, only when they find less than 5 similar sequences under the specific conditions of the BLAST program will they be considered specific and adopted.

[0113] Polymerase chain reaction (PCR) was performed using the AmpliTaq(R) Gold polymerase kit (Applied ...

Embodiment 2

[0117] Example 2: Detection of the newly discovered single nucleotide polymorphism site 67044_67045delT in this invention

[0118] In this invention, a single nucleotide polymorphism site 67044_67045delT located on exon 16 of the ABCG2 gene was newly discovered. The method for detecting this site is introduced as follows:

[0119] With the software Primer3 ( http: / / frodo.wi.mit.edu / cgi-bin / primer3 / primer3 ) Design some specific primers to search for polymorphic sites in related regions. use BLAST TM The program (National Center for Biotechnology Information - http: / / www.ncbi.nlm.nih.gov / BLAST) tests the specificity of the designed primers relative to the human genome sequence. Whether forward or reverse primers, only when they find less than 5 similar sequences under the specific conditions of the BLAST program, they are considered specific and adopted.

[0120]Polymerase chain reaction (PCR) was performed with AmpliTaq(R) Gold polymerase kit (Applied Biosystems, CA, USA) ...

Embodiment 3

[0124] Example 3: Detection of SNPs associated with colorectal cancer on the human ABCG2 gene

[0125] This example describes a method for detecting and identifying one or several of the SNPs described above. A partial target region of the human ABCG2 gene will be used as a template to synthesize PCR products. Table 4 lists several pairs of specific primers, each pair can be used to amplify a specific target region.

[0126] single nucleotide polymorphism

[0127] PCR Amplification of Fragments Containing Single Nucleotide Polymorphisms and Sequencing of Amplified Products

[0128] The following reagents were used for PCR amplification of the fragment of interest: 1.5 mM magnesium ions, 200 μM of the four bases, 0.3 μM for each primer, 10 ng of template genomic DNA, and 0.5 U of polymerase. The total volume of the polymerase chain reaction was 20 µl.

[0129] The polymerase chain reaction was performed using a Mastercycler(R) Gradient thermal cycler (Eppendorf, Ha...

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Abstract

The present invention relates to the detection of 15 mononucleotide polymorphism sites related with colorectal carcinoma and on 6 heterogenous substance metabolizing enzyme gene, the application of these mononucleotide polymorphism sites in predicting the liability of colorectal carcinoma, and the method of diagnosing and treating colorectal carcinoma. The present invention provides information favorable to preventing colorectal carcinoma as one main deadly disease and developing new effective therapy.

Description

technical field [0001] The present invention relates to the detection of 15 single nucleotide polymorphism (SNP) sites associated with colorectal carcinoma (CRC) on 6 heterologous substance metabolizing enzyme genes, and the application of these single nucleotide polymorphisms Sexual loci predict susceptibility to colorectal cancer, and approaches to diagnosis and treatment of this common cancer. The present invention provides a method that helps prevent colorectal cancer and develops an effective new therapy. The present invention can also be applied to screen for appropriate therapy against different types of colorectal cancer. Background technique [0002] Colorectal cancer is the second most common malignancy and the second leading cause of cancer death in Western countries. In the Caucasian population, the incidence of colorectal cancer approaches that of lung cancer (see Levi F, Lucchini F, Negri E., et al 1999. Cancer mortality in Europe, 1990-1994 and overview of t...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07H21/04C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 吴汪黔生张浩廖凌虹
Owner PHARMACOGENETICS
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