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Method for detecting gastric polyp and gastric cancer using marker gene of gastric polyp and gastric cancer-specific methylation

A technology for methylation and gastric polyps, which is applied in the fields of biochemical equipment and methods, measuring devices, and microbial determination/inspection, and can solve problems such as no suggestion.

Inactive Publication Date: 2015-07-29
GENOMICTREE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the literature does not suggest the use of the Ligandrecan 2 gene for the diagnosis of other cancers including gastric cancer

Method used

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  • Method for detecting gastric polyp and gastric cancer using marker gene of gastric polyp and gastric cancer-specific methylation
  • Method for detecting gastric polyp and gastric cancer using marker gene of gastric polyp and gastric cancer-specific methylation
  • Method for detecting gastric polyp and gastric cancer using marker gene of gastric polyp and gastric cancer-specific methylation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0143] Example 1: Measurement of methylation of the SDC2 biomarker gene in tissues of patients with gastric polyps

[0144] To evaluate the usefulness of the SDC2 biomarker gene for the early diagnosis of precancerous gastric polyps, paraffin tissues from normal gastric tissues (Biochain; 5 samples) and patients with gastric polyps (provided by Chungnam National University Hospital's biobank; 10 samples with low-grade dysplasia and 10 samples with high-grade dysplasia) to isolate genomic DNA. Isolation of genomic DNA from paraffin tissue was performed using the QIAamp DNA Micro Kit (Qiagen, Germany) according to the manufacturer's instructions.

[0145] Methylation assays were performed using a quantitative pyrosequencing method. 200 ng of isolated genomic DNA was treated with bisulfite using the EZ DNA Methylation-Gold Kit (Zymo Research, USA). When DNA is treated with bisulfite, unmethylated cytosines are modified to uracils, while methylated cytosines remain unchanged. B...

Embodiment 2

[0156] Example 2: Measurement of Methylation of Biomarker Genes in Gastric Cancer Cell Lines and Gastric Cancer Tissues

[0157] In order to examine whether the SDC2 biomarker gene confirmed to be methylated in gastric polyps can also be used as a biomarker for diagnosing gastric cancer, pyrosequencing was performed in the same manner as described in Example 1.

[0158] Using the QIAamp DNA Mini Kit (Qiagen, Germany) according to the manufacturer's instructions, the gastric cancer cell line AGS (Korean Cell Line Bank (KCLB No.21739) and 41 gastric cancer patients' cancer tissues and adjacent normal tissues (provided by the Biobank of Chungnam National University Hospital) isolated genomic DNA. Using the EZ DNA methylation-gold kit (Zymo Research, USA), the genomic DNA of 200ng isolation was treated with bisulfite. When the genomic DNA was treated with sulfurous acid Upon hydrogen salt treatment, unmethylated cytosines were modified to uracil while methylated cytosines remained...

Embodiment 3

[0164] Example 3: Measurement of methylation of SDC2 biomarker gene in serum of patients with gastric cancer

[0165] In order to examine the usefulness of the SDC2 biomarker gene as a biomarker for gastric cancer diagnosis using serum, the methylation of the SDC2 biomarker gene in the serum of gastric cancer patients was measured by the quantitative methylation-specific real-time PCR (qMSP) method.

[0166] For this purpose, two PCR primers (IDT, USA) and a fluorescent probe (IDT, USA) capable of specifically amplifying bisulfite-treated methylated SDC2 gene were designed. To determine the amount and quality of bisulfite-treated serum DNA, the ACTB gene was used as an internal control. The sequences of PCR primers and fluorescent probes used in qMSP are shown in Table 2 below.

[0167] Table 2

[0168] [Table 2]

[0169] Primer and fluorescent probe sequences for qMSP

[0170]

[0171] Genomic DNA was isolated from 800 μl serum using the DynalBeads SILANE viral NA kit ...

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Abstract

The present invention relates to a novel use of the SDC2 (NM_002998, syndecan 2) gene as a biomarker of a gastric polyp and gastric cancer-specific methylation, and more particularly, to a use for diagnosing a gastric polyp and gastric cancer in an early stage by measuring the methylation level using the SDC2 gene as a biomarker. The present invention has the effect of providing a method which can provide information for the diagnosis of gastric cancer by detecting the methylation of a CpG island of a gastric polyp and a gastric cancer-specific marker gene. Also, using the method for detecting methylation and a composition, kit, and nucleic acid chip for diagnosis according to the present invention, gastric cancer can be diagnosed at an initial transformation stage, and thus, early diagnosis can be achieved. Therefore, the present invention is useful as it is possible to diagnose gastric cancer more accurately and rapidly than conventional methods.

Description

technical field [0001] The present invention relates to the novel use of the syndecan-2 (syndecan, SDC2; NM_002998) gene as a methylation biomarker specific for gastric polyps and gastric cancer, and more particularly to the syndecan-2 gene Use as a biomarker that enables the diagnosis of gastric polyps and gastric cancer at an early stage by measuring the methylation level of the syndecan-2 gene. Background technique [0002] Even with the development of medicine today, the 5-year survival rate of cancer patients, especially those with solid tumors (except blood cancer patients), is still less than 50%, and about 2 / 3 of all cancer patients are diagnosed at an advanced stage, and almost all of them are diagnosed with cancer Died within 2 years of diagnosis. Such poor outcomes in cancer treatment are not only a matter of treatment methods, but also due to the fact that it is not easy to diagnose cancer at an early stage and accurately diagnose cancer at an advanced stage, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6886C12Q2600/154G01N33/57446
Inventor 安城皖吴泰祯
Owner GENOMICTREE
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