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miRNA-6761-5p and new application thereof

A technology of colon cancer and diagnostic reagents, applied in the field of molecular biology, can solve the problems of no research showing the relationship between digestive tract tumors and few correlations

Pending Publication Date: 2019-05-21
天津市中西医结合医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that miRNA plays a very important role in the occurrence and development of colon cancer: miR-720 and miR-484 in serum can be used for staging and early diagnosis of colon cancer; miR-93 may participate in colon cancer through the PI3K / Akt signaling pathway The process of occurrence and development; patents ZL2014101918129, CN2016106181508, and CN201610620059X all reveal the correlation between miRNA and colon cancer, but currently few studies focus on the correlation between miRNA and left and right colon cancer
[0004] In this study, the applicant integrated the high-throughput sequencing data in the TCGA database and analyzed it using bioinformatics methods, and found that miR-6761-5p, a potential target that can distinguish left-sided colon cancer from right-sided colon cancer, was reported in existing research miR-6761-5p is associated with melanoma, but no studies have shown its relationship with gastrointestinal tumors

Method used

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  • miRNA-6761-5p and new application thereof
  • miRNA-6761-5p and new application thereof
  • miRNA-6761-5p and new application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Distinguishes the screening of miRNAs associated with left and right colon adenocarcinomas

[0042] 1. Data collection

[0043] At the time of retrieval, the TCGA database included 460 patients with colon adenocarcinoma, including clinical data of 459 patients, mRNA data of 456 patients, and miRNA data of 444 patients.

[0044] According to the clinical data records, the patients included in the study were screened. Require:

[0045] (1) histological_Type is colon adenocarcinoma;

[0046] (2) anatomic_neoplasm_subdivision is Ascending Colon, Sigmoid Colon, Cecum, Descending Colon.

[0047] A total of 314 patients were screened out, among which the adenocarcinoma of the right colon included: ascending colon, cecum (Cecum); the adenocarcinoma of the left colon included: descending colon, sigmoid colon.

[0048] Colon adenocarcinoma transcriptome mRNA and miRNA data generated by TCGA database were used. After screening according to the above criteria, the sa...

Embodiment 2

[0053] Example 2 Q-PCR verification of differentially expressed and miR-6761-5p

[0054] 1. Blood samples from 42 patients with colon cancer were collected from the Department of Gastrointestinal Surgery, Nankai Hospital, Tianjin, including 20 right colon cancer samples and 22 left colon cancer samples. All samples were pathologically confirmed.

[0055] 2. RNA extraction

[0056] Use the QIAGEN blood RNA extraction kit to extract RNA in serum, and refer to the instruction manual for specific steps.

[0057] 3. Reverse transcription:

[0058] 1) Mix 10pg-1μg total RNA template with 2μl 10× buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl RNase inhibitor and RNase free water , the final volume was 20 μl, and incubated at 37°C for 1h.

[0059] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0060] 3) Immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers...

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Abstract

The invention relates to miRNA-6761-5p and new application thereof, in particular to miRNA-6761-5p and application thereof to preparation of a diagnosis preparation for distinguishing a left-sided colon cancer from a right-sided colon cancer. Left-sided and right-sided colons are different in source and have difference in the aspects of physiological functions, forms, metabolism and others, so that tumors have large difference in pathogenesis. Through analysis of high-flux data, it is found that miRNA-6761-5p has remarkable differential expression in the left-sided colon cancer, then a fluorescent quantitative PCR experiment is used for verifying and analyzing, and results show that miRNA-6761-5p is possible to be a molecular marker for distinguishing the left-sided colon cancer from the right-sided colon cancer, and lays foundations for clinical application.

Description

technical field [0001] The present invention relates to the field of molecular biology, and more specifically relates to miR-6761-5p and its application in preparing diagnostic preparations for distinguishing left colon cancer and right colon cancer. Background technique [0002] The colon is divided into the right colon and the left colon, which are bounded by the splenic flexure. The former includes the ileocecal, ascending colon, hepatic flexure, and transverse colon; the latter consists of the splenic flexure, descending colon, and sigmoid colon. The 2017 edition of the NCCN guidelines clarifies the mainstream view of the medical community on left and right colon cancer, that is, the embryonic origin of the left and right colons is different, and there are differences in physiological functions, morphology, and metabolism between the two, which lead to tumors in pathogenesis, pathological characteristics, and clinical manifestations. The characteristics, metastasis rules...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
Inventor 于向阳饶阳王西墨敖奇邹常林杨士民李琳王彤彤
Owner 天津市中西医结合医院
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