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Early molecular diagnosis marker for esophageal squamous cell carcinoma and application of marker

A technology for esophageal squamous cell carcinoma and esophageal cancer, applied in the direction of medical preparations containing active ingredients, drug combinations, organic active ingredients, etc., can solve the problems of unclear diagnosis and lost treatment opportunities for patients with esophageal squamous cell carcinoma

Active Publication Date: 2017-12-29
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the unclear diagnosis caused by the level of technology, it is likely that some patients with early esophageal squamous cell carcinoma will lose the best treatment opportunities

Method used

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  • Early molecular diagnosis marker for esophageal squamous cell carcinoma and application of marker
  • Early molecular diagnosis marker for esophageal squamous cell carcinoma and application of marker
  • Early molecular diagnosis marker for esophageal squamous cell carcinoma and application of marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The collection of embodiment 1 sample

[0050] The samples of 3 cases of metastatic esophageal squamous cell carcinoma tissue, 3 cases of non-metastasis esophageal squamous cell carcinoma tissue and 6 cases of paracancerous tissue were obtained from surgical resection specimens in the hospital from June 2015 to January 2017. The body was placed in a liquid nitrogen tank within 10 minutes, and then transferred to a -80°C refrigerator for storage.

Embodiment 2

[0051] Example 2 total RNA extraction

[0052] 1 extraction method

[0053] 1) Take 80 mg tissue block, add 800 μl Lysis / Binding buffer, and use a homogenizer to homogenize the tissue block. During the homogenization process, the samples should be placed on ice to keep the cold state.

[0054] 2) Add 1 / 10 volume of Homogenate Additive to the above homogenized tissue samples, and place on ice for 10 minutes.

[0055] 3) Add an equal volume of water-saturated phenol to the Lysis / Binding buffer, shake for 45 seconds, and centrifuge at 10,000×g for 5 minutes at room temperature.

[0056] 4) Carefully remove the supernatant into a new test tube, add 1.25 times the volume of absolute ethanol, mix well, transfer to a purification column, centrifuge at 10,000×g for 15 s, and discard the liquid in the collection tube. Since the maximum volume of the column is only 700 μl, repeat this step until all the supernatant is filtered.

[0057] 5) Add 700 μl of miRNA eluent 1 to the spin co...

Embodiment 3

[0066] Example 3 Sequencing and Data Analysis

[0067] The sequencing company carried out the establishment of the sequencing library and on-machine sequencing, and the sequencer used was the HiSeq2000 sequencer of Illumina Company.

[0068] Statistical analysis was performed according to the data provided by the sequencing company, fdr1, and the difference between the mean count values ​​of the two groups was greater than 100. Differentially expressed miRNAs were artificially selected to filter miRNAs that were significantly differentially expressed, and several previously unreported molecular markers related to esophageal squamous cell carcinoma entered our research scope, among which miR-1277-5p was in the metastasis group and the diseased group. Although there was differential expression between the two groups, the difference was not significant, and the expression levels in metastatic and diseased cancer tissues were significantly higher than those in paracancerous tissue...

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Abstract

The invention relates to an early molecular diagnosis marker for esophageal squamous cell carcinoma and an application of the marker. Parts of patients suffering from early esophageal squamous carcinoma may lose the best treatment opportunity due to indefinite diagnosis caused by the existing diagnostic technique level of the esophageal squamous cell carcinoma, more accurate indexes are required for supplement, and a molecular biology method for researching and identifying effective molecule markers is a key means for assisting existing clinical diagnosis, clinical intervention guide and precancerous warning. The provided early molecular diagnosis marker mir-1277 for esophageal squamous cell carcinoma and mature miRNA provide a potential basis for accurate diagnosis of clinical esophageal squamous cell carcinoma and have important actual application value.

Description

technical field [0001] The present invention relates to the field of gene detection, in particular to early molecular diagnostic markers of esophageal squamous cell carcinoma and its application in clinical diagnosis and treatment, and more specifically to the application of mir-1277 and its mature miRNA in the preparation of diagnostic preparations for esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer is one of the top ten common malignant tumors in humans. my country is one of the countries with a high incidence of esophageal cancer, especially esophageal squamous cell carcinoma (ESCC) is the main pathological type of esophageal cancer in my country. The occurrence of esophageal squamous cell carcinoma experienced a process from normal esophageal mucosa, inflammation, esophageal epithelial dysplasia, carcinoma in situ, intramucosal carcinoma, early invasive carcinoma, and advanced carcinoma. The onset of esophageal cancer is hidden, and ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61P35/00A61P1/00C12Q1/68
CPCA61K31/7105C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 任静宋宏涛石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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