Novel application of mir-3613 and mature miRNA thereof

A mature technology of mir-3613, applied in the field of genetic testing, can solve problems such as unsatisfactory prognosis, low survival rate, and poor prognosis

Inactive Publication Date: 2017-12-05
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current treatment of esophageal squamous cell carcinoma is based on radical resection combined with perioperative chemotherapy or chemoradiotherapy, but the disease progresses rapidly, the prognosis is poor, and the long-term efficacy is still unsatisfactory. The 5-year overall survival rate is very low, and the prognosis effect is not good. ideal
There is no ideal and reliable diagnostic marker for esophageal squamous cell carcinoma

Method used

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  • Novel application of mir-3613 and mature miRNA thereof
  • Novel application of mir-3613 and mature miRNA thereof
  • Novel application of mir-3613 and mature miRNA thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The collection of embodiment 1 sample

[0053] 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

[0054] Example 2 total RNA extraction

[0055] 1 extraction method

[0056] 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.

[0057] 2) Add 1 / 10 volume of Homogenate Additive to the above homogenized tissue samples, and place on ice for 10 minutes.

[0058] 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.

[0059] 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.

[0060] 5) Add 700 μl of miRNA eluent 1 to the spin co...

Embodiment 3

[0069] Example 3 Sequencing and Data Analysis

[0070] 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.

[0071] 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-3613-5p 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 a novel application of mir-3613 and mature miRNA thereof. Based upon transcriptome sequencing on samples of metastatic esophageal squamous carcinoma tissues, non-metastatic esophageal squamous carcinoma tissues and para-carcinoma tissues, as well as analysis which is implemented in accordance with a bio-informatics method, some molecular markers related to esophageal squamous carcinoma are discovered, and the expression quantity of miR-3613-5p in the carcinoma tissues is higher than that of miR-3613-5p in the para-carcinoma tissues; and furthermore, a fluorescent quantitative PCR experimental result is consistent with a high-throughput sequencing result. The invention offers potential molecular markers for the precise diagnosis of the esophageal squamous carcinoma in the clinical field; and an important practical application value is achieved.

Description

technical field [0001] The present invention relates to the field of gene detection, in particular to miRNA related to esophageal squamous cell carcinoma and its application, and more specifically to the application of mir-3613 and its mature miRNA in preparing diagnostic preparations for esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer is a common malignant tumor of the digestive tract. About 300,000 people die from esophageal cancer every year, and its morbidity and mortality rank among the top few malignant tumors. my country is one of the countries with high incidence of esophageal cancer, especially esophageal squamous cell carcinoma (ESCC) is the main pathological type of esophageal cancer in my country. The current treatment of esophageal squamous cell carcinoma is based on radical resection combined with perioperative chemotherapy or chemoradiotherapy, but the disease progresses rapidly, the prognosis is poor, and the long-term effi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K31/7105A61K48/00A61P35/00
CPCA61K31/7105C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 石小峰任静宋宏涛
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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