Colorectal cancer diagnostic biomarker and application thereof

A biomarker, colorectal cancer technology, applied in biochemical equipment and methods, microbial determination/inspection, organic compound library, etc. problems such as high misdiagnosis rate, to achieve the effect of improving 5-year survival rate, broad application prospects and timely diagnosis

Active Publication Date: 2017-12-08
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above detection methods still have certain limitations, such as the cumbersome separation process of fecal occult blood examination, which is easi

Method used

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  • Colorectal cancer diagnostic biomarker and application thereof
  • Colorectal cancer diagnostic biomarker and application thereof
  • Colorectal cancer diagnostic biomarker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: High-throughput chip processing and analysis of circular RNA

[0054] Firstly, the raw data is filtered, and then the data with low-quality signal and noise pollution are removed, and the effective circular RNA expression value is obtained after normalization. On the basis of the above analysis, a series of statistical and visual analysis can be carried out.

[0055] In this study, the multiple of change (Fold Change, FC) ≥ 1.5 and P figure 2 ). Then the screened differentially expressed mRNAs were represented by a heat map ( image 3 ).

Embodiment 2

[0056] Example 2: Detection of differential expression of hsa_circ_0006110 gene in colorectal cancer cell lines and normal intestinal epithelial cell lines by fluorescent quantitative PCR

[0057] 1. Cultivate human colorectal cancer cell lines (SW620, HCT116 and RKO) and human normal intestinal epithelial cell line NCM460. Each 100U / mL of DMEM medium at 37°C and 5% CO 2 Cultured in an incubator, the culture medium was changed every 2-3 days and subcultured. Human intestinal cancer HCT116, SW620 and RKO cell lines were treated with DMEM (containing 10% fetal bovine serum) at 37°C and 5% CO 2 cultured in an incubator.

[0058] 2. When the confluence of the cells in the 60mm culture dish reaches 80%, discard the culture medium, add 1mL sterile PBS to wash, discard the PBS and repeat once, add 1mL Trizol RNA extraction solution, and gently shake the culture dish left and right , so that all the Trizol RNA extraction solution was submerged at the bottom of the culture dish, gen...

Embodiment 3

[0076] Example 3: RT-PCR reaction detection of differential expression of hsa_circ_0006110 gene in colorectal cancer and paracancerous tissues.

[0077] 1. Collect 25 pairs of colorectal cancer tissue and corresponding distant normal intestinal tissue samples. Each patient sample has a clear diagnosis record, and with the consent of the ethics committee, the samples are collected and frozen in liquid nitrogen.

[0078] 2. Preparation of RNA samples

[0079] (1) Pretreatment of the sample: Take the sample out of the liquid nitrogen tank, put the sample into a 2mL sterile EP tube, add 1mL Trizol RNA extraction solution, soak it in 0.1% DEPC water overnight and cut it into pieces with sterilized scissors. Grind thoroughly in a tissue homogenizer.

[0080] 3. RNA extraction, reverse transcription, fluorescent quantitative PCR and data analysis methods are the same as in Example 2.

[0081] 4. Results: The expression level of hsa_circ_0006110 was detected in 25 pairs of colorecta...

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Abstract

The invention relates to a colorectal cancer diagnostic biomarker and application thereof. A nucleotide sequence is shown as SEQ ID NO. 1. The colorectal cancer diagnostic biomarker provided by the invention has the beneficial effects that hsa_circ_0006110 gene expression is discovered to be closely related to a colorectal cancer for the first time; through detecting the expression of hsa_circ_0006110 in a subject colorectal tissue, whether a subject suffers from the colorectal cancer or the risk of the colorectal cancer exists or not can be more accurately and quickly judged, so that a prevention or treatment scheme is provided for a clinician. A target gene as a drug for treating the colorectal cancer provides a new treatment targets and treatment approach for colorectal cancer treatment; compared with a traditional detection means, the diagnostic biomarker is more timely and more specific to diagnose, so that the five-year survival rate of a colorectal cancer patient is improved, and the death rate is reduced, therefore, the application prospect is wide.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a circular RNA hsa_circ_0006110 and its application as a marker in the diagnosis of colorectal cancer. Background technique [0002] Colorectal cancer (CRC) is one of the most common malignant tumors of the digestive tract, the third most common tumor in men and the second in women, and the second leading cause of cancer deaths. According to the International Agency for Research on Cancer of the World Health Organization, there were about 1.2 million new cases of colorectal cancer in the world in 2008, and its mortality rate accounted for about 8% of all malignant tumors. With the improvement of my country's economy and life in the past 30 years, the incidence and death of colorectal cancer in my country are on the rise, ranking third in the incidence of malignant tumors and fifth in death in my country, seriously threatening people's health and life . Colo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C40B40/06
CPCC12Q1/6837C12Q1/6886C12Q2600/158C12Q2600/178C40B40/06
Inventor 陈瑞李晓波陆润泽孟庆涛吴申申孙浩
Owner SOUTHEAST UNIV
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