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Internal reference miRNA as esophagus cancer sample and application of internal reference miRNA

A technology for esophageal cancer and samples, which is applied in the field of miRNA detection, can solve the problems of miRNA difficulties, lack of internal reference genes, and limitations of miRNA research and application, and achieve good stability, consistent expression abundance, and high serum expression abundance.

Active Publication Date: 2019-05-31
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, although some progress has been made, the research and application of miRNA in esophageal cancer are still limited.
Every miRNA researcher must face a very thorny problem when detecting healthy subjects and esophageal cancer-related patients, the lack of reliable internal reference genes makes it difficult to compare miRNAs between samples
In addition, the current research on miRNA in esophageal cancer is mainly focused on the Caucasian population in Europe and the United States, and there is a lack of miRNA internal references suitable for subspecies, especially the Chinese population.

Method used

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  • Internal reference miRNA as esophagus cancer sample and application of internal reference miRNA
  • Internal reference miRNA as esophagus cancer sample and application of internal reference miRNA
  • Internal reference miRNA as esophagus cancer sample and application of internal reference miRNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This embodiment aims at preliminary screening to obtain candidate esophageal cancer internal reference miRNAs, and the specific screening methods and results are as follows:

[0033] (1) Select the following seven miRNAs as candidate internal reference miRNAs for esophageal cancer: hsa-miR-361-5p, hsa-miR-186-5p, hsa-miR-26a-5p, hsa-miR-191-5p, hsa- miR-451a, hsa-miR-423-5p and hsa-miR-320a.

[0034] (2) With the help of the data source of TCGA (The Cancer Genome Atlas) database, the expression levels of the aforementioned 7 miRNAs in esophageal cancer tissues and their adjacent tissues were investigated. No significant difference.

[0035] (3) With the help of the data source of the miRNA-seq database, the expression levels of the aforementioned seven miRNAs in serum were investigated, and the results showed that the abundance of hsa-miR-361-5p and hsa-miR-186-5p in serum Low, not suitable as internal reference miRNA.

[0036] (4) According to literature review, hsa...

Embodiment 2

[0039] The purpose of this example is to investigate the expression abundance of hsa-miR-423-5p, hsa-miR-451a and hsa-miR-320a in the samples of normal subjects and esophageal cancer subjects in the Chinese population, in order to investigate their Whether it is suitable as an internal reference miRNA for esophageal cancer. Among them, the subjects consisted of 32 normal subjects and 32 esophageal cancer subjects, all selected from the Chinese population; the test samples were serum, and the detection method was qRT-PCR. The specific detection steps are as follows:

[0040] (1) RNA extraction

[0041] Using miRNeasy Serum / Plasma kit (Qiagen) to extract serum RNA, the operation steps are as follows: 250 μl serum samples were centrifuged (10,000 RPM, 5mins) to remove cell debris, 200 μl supernatant was transferred to a new RNase-removing centrifuge tube; 1,000 μl of The QIAzol lysate was finally dissolved in 30 μl RNase-removed water according to the operation steps provided i...

Embodiment 3

[0059] This example aims to expand the number of samples and further verify the expression level of hsa-miR-423-5p in samples from normal subjects and esophageal cancer subjects. Among them, the subjects consisted of 100 normal subjects and 100 esophageal cancer subjects, all selected from the Chinese population; the test samples were serum, and the detection method was qRT-PCR. Refer to Example 2 for specific detection steps. For specific test results, see Figure 4 .

[0060] according to Figure 4 The results shown show that the expression levels of hsa-miR-423-5p in the serum of normal subjects and esophageal cancer subjects are basically the same without significant difference (P=0.18), and the expression abundance is high, which is suitable as a Internal reference genes of miRNA in esophageal cancer.

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Abstract

The invention relates to an internal reference miRNA as an esophagus cancer sample and application of the internal reference miRNA. The internal reference miRNA as the esophagus cancer sample includeshsa-miR-423-5p and / or hsa-miR-16-5p, wherein hsa-miR-423-5p is high in expression richness and low in fluctuation amplitude when expressed in the sample, has no obvious differences among different types of samples and is suitable as the esophagus cancer sample, especially as the internal reference miRNA as the esophagus cancer sample for Chinese people.

Description

technical field [0001] The invention relates to the field of miRNA detection, in particular to an internal reference miRNA of esophageal cancer samples and its application. Background technique [0002] Esophageal cancer (esophageal carcinoma, EC) is one of the most frequent malignant tumors of the digestive tract in the world. About 400,000 people die from this disease every year in the world, seriously endangering public health. The incidence of esophageal cancer has obvious regional differences. It is one of the areas with high incidence of upper esophageal cancer, with an average incidence rate of about 22.14 / 100,000 and a mortality rate of 16.77 / 100,000 during the same period, ranking fourth in the mortality rate of malignant tumors. Esophageal cancer has the characteristics of resistance to radiotherapy and chemotherapy, local recurrence or distant metastasis, and the five-year survival rate is about 15-25%. The early symptoms of esophageal cancer are mostly atypical,...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
Inventor 刘芝华高峰骆爱萍周宣彤王慜杰
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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