Methylation site in the promoter region of eno1 gene and its application in the preparation of kits for early diagnosis and prognosis evaluation of esophageal cancer

A gene promoter region and early diagnosis technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problem of lower overall survival rate of patients, and achieve far-reaching clinical significance and promotion value. Good specificity and easy operation

Active Publication Date: 2022-06-24
NANJING DRUM TOWER HOSPITAL
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Problems solved by technology

Another study reported that patients with high expression of ENO1 had a lower overall survival rate

Method used

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  • Methylation site in the promoter region of eno1 gene and its application in the preparation of kits for early diagnosis and prognosis evaluation of esophageal cancer
  • Methylation site in the promoter region of eno1 gene and its application in the preparation of kits for early diagnosis and prognosis evaluation of esophageal cancer
  • Methylation site in the promoter region of eno1 gene and its application in the preparation of kits for early diagnosis and prognosis evaluation of esophageal cancer

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Experimental program
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Effect test

Embodiment 1

[0071] Methylation changes of ENO1 gene in esophageal squamous cell carcinoma and its clinicopathological significance

[0072] 1. DNA preparation: The tumor tissues of 68 patients with esophageal squamous cell carcinoma and their corresponding 62 adjacent normal tissues were selected, fixed in formalin and embedded in paraffin. The paracancerous normal tissues were all paraffin blocks of esophageal normal tissues and were selected from samples at a distance of more than 2 cm from the esophageal squamous cell carcinoma. Genomic DNA was extracted with QIAamp DNA FFPE Tissue Kit, and its content and purity were determined by UV spectrophotometer to determine the absorbance value, and gel electrophoresis was used to ensure the quality of the extracted DNA.

[0073] 2. Bisulfite treatment: Use EZ-96DNA Methylation-Gold™ Kit for bisulfite modification. The obtained DNA was used immediately or stored at -20°C for later use.

[0074] 3. PCR amplification reaction: Design primers fo...

Embodiment 2

[0129] Expression of ENO1 protein in esophageal squamous cell carcinoma tissue and adjacent normal.

[0130] The expression of ENO1 protein in 273 cases of esophageal squamous cell carcinoma and 263 cases of adjacent normal tissues was detected by immunohistochemistry.

[0131] The prepared esophageal cancer tissue chip paraffin block was taken for 4 μm serial sections, and the immunohistochemical method of the tissue chip was the same as the conventional method. This experiment adopts the two-step immunohistochemical Envision method, and the specific steps are as follows:

[0132] (1) Before the experiment, put the prepared 4μm white slices into the slicing box and store them in a 4°C refrigerator for subsequent experiments;

[0133] (2) Before the experiment, insert the white slices of the tissue chips stored in the refrigerator at 4°C into the metal basket and bake them in a 60°C oven for 1.5-2 hours to prevent the tissue from falling off;

[0134] (3) Put the paraffin ti...

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Abstract

The invention provides the methylation site of the ENO1 gene promoter region and its application in the preparation of a kit for early diagnosis and prognosis evaluation of esophageal cancer, belonging to the technical field of disease diagnosis and prognosis rating; the methylation site of the ENO1 gene promoter region As shown in SEQ ID No.1, the DNA of esophageal squamous cell carcinoma and adjacent normal samples was extracted, modified with bisulfite, and the methylation frequency of the CpG site in the promoter region of the ENO1 gene was detected by flight mass spectrometry. Immunohistochemical detection of ENO1 protein expression in esophageal squamous cell carcinoma and adjacent normal tissue samples; the study found that ENO1 gene promoter region was hypomethylated and ENO1 protein was highly expressed in esophageal squamous cell carcinoma, and patients with low methylation had poor prognosis , ENO1 can promote the glycolysis level of esophageal cancer cells, promote its proliferation and inhibit apoptosis; detect the methylation level of ENO1 gene for early diagnosis and prognosis evaluation of esophageal squamous cell carcinoma.

Description

technical field [0001] The invention belongs to the technical field of disease diagnosis and prognosis rating, and particularly relates to a methylation site in the promoter region of ENO1 gene and its application in preparing a kit for early diagnosis and prognosis evaluation of esophageal cancer. Background technique [0002] Esophageal carcinoma (EC) is the sixth leading cause of death-related cancer worldwide. Esophageal squamous cell carcinoma (ESCC) is the most common histological subtype of esophageal cancer, especially in East Asia, eastern and southern Africa. In China, about 90% of esophageal cancer patients are esophageal squamous cell carcinoma. Esophageal cancer in China has obvious geographical distribution, and the incidence of esophageal cancer ranks first among all tumors in Henan, Hebei, Jiangsu, Shaanxi, Shanxi, Fujian and other places. Despite improvements in diagnosis and treatment, the survival rate of patients with advanced esophageal squamous cell c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154C12Q2600/158C12Q2600/118
Inventor 崔晓宾赵琳玥王文洁陈云昭孙琦丁友祥禹洁陈曦李锋韩雪萍孟凡青
Owner NANJING DRUM TOWER HOSPITAL
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