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miRNA related to colorectal cancer and application thereof

A technology for colon cancer and rectal cancer, applied in miRNA and its application fields, can solve the problems of easy to miss micrometastases, inaccurate tumor staging, time-consuming, etc., and achieve the effect of improving healing power, reducing patient mortality, and low cost

Pending Publication Date: 2020-12-11
GUANGZHOU FULENGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The former depends on the judgment of histopathologists. This method is not only time-consuming for initial diagnosis, but also easily misses micrometastasis in lymph node histological examination, resulting in inaccurate staging of resected tumors; the safety and examination cost of the latter are also worth considering

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Screening method for miRNAs associated with colon cancer and rectal cancer

[0025] (1) Establish a standardized and normalized colon cancer and rectal cancer tissue and body fluid specimen bank and case database in Guangzhou

[0026] The Third Affiliated Hospital of Guangzhou Medical University (referred to as "the Third Hospital of Guangzhou Medical University") is the participating unit of the project. The establishment of the specimen bank and database was established by the Third Hospital of Guangzhou Medical University. More than 200 cases of tumors, Adjacent tissue and plasma were stored at -80°C, and the collected clinical data (such as hospitalization number, pathology number, tissue location, tumor stage and grade, surgical method, chemotherapy regimen, follow-up) and experimental data (such as : Extracted miRNA concentration, spectrophotometry, electrophoresis pictures, quantitative PCR experiment results) were counted and organized, and a colon ca...

Embodiment 2

[0039] Example 2: Differences in the expression of miR-1290 in the plasma of patients with colon cancer and rectal cancer and healthy persons

[0040] Fluorescent quantitative PCR was used to detect the expression of miR-1290 in the plasma of 219 patients with colon cancer and rectal cancer and 45 age- and sex-matched healthy subjects, and to analyze the relationship between it and clinical characteristics. The specific methods are as follows:

[0041] (1) Collection of plasma samples: 2 mL of peripheral blood from each subject was collected in EDTA test tubes. After standing still, the supernatant was collected by centrifugation, and stored in a -80°C refrigerator for later use.

[0042] (2) Extraction of plasma total RNA: RNA extracted by TRIzol method was stored in a -80°C refrigerator for later use.

[0043] (3) cDNA synthesis: Using 2 μg of total RNA as a template, prepare a reverse transcription reaction system according to the Bestar qPCR RT Kit instructions, synthesize...

Embodiment 3

[0049] Example 3: Real-time fluorescent quantitative PCR detection kit for colon cancer and rectal cancer based on miR-1290

[0050] This embodiment provides a kit for distinguishing colon cancer samples and rectal cancer samples from normal samples in blood samples of subjects, the kit includes: a detection reagent suitable for detecting miR-1290; A device for determining the amount of miR-1290 detected by the detection reagent.

[0051] The kit is a real-time fluorescent quantitative PCR detection kit for colon cancer and rectal cancer, including a test strip with an oligonucleotide probe, the sequence of which is the complementary sequence of miR-1290.

[0052] Other reagents of this kit can be prepared with reference to known real-time fluorescent quantitative PCR detection kits.

[0053] The detection steps of the kit include: (1) extraction of total RNA of the sample; (2) detection of RNA quality; (3) reverse transcription of RNA; (4) real-time fluorescent quantitative ...

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Abstract

The invention belongs to the field of nucleic acid diagnosis and treatment, and particularly relates to miRNA related to colorectal cancer and application thereof. The miRNA related to colorectal cancer is miR-1290. The invention provides application of the miR-1290 in preparing a kit for distinguishing a colorectal cancer sample and a rectal cancer sample from a normal sample in a blood sample ofa subject. The invention determines that the miR-1290 is a miRNA diagnostic marker related to colorectal cancer for the first time, and establishes a novel colorectal cancer and rectal cancer tumor progression detection index. On the basis, the invention provides novel application of the miR-1290 in the aspect of initial diagnosis and rapid detection of colorectal cancer and rectal cancer and a related universal applicable kit.

Description

technical field [0001] The invention belongs to the field of nucleic acid diagnosis and treatment, and in particular relates to a miRNA related to colon cancer and rectal cancer and its application. Background technique [0002] Colon cancer and rectal cancer are one of the most common malignant tumors that seriously endanger people's health. They rank first in cancer incidence and second in mortality in developed countries, and fourth in cancer mortality in my country, and the incidence is increasing year by year. In Guangzhou, the crude incidence and mortality rates of colon cancer and rectal cancer were 9.6 / 100,000 and 5.0 / 100,000, respectively, and increased by nearly 5%. The survival and prognosis of patients depend on the detection time of the tumor. Early detection and early treatment are the only ways to improve the long-term survival rate of colon cancer and rectal cancer. For example, according to Dukes staging, the 5-year survival rate can reach 90±% in stage A ,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6837C12Q1/6834
Inventor 李华平张金阔吴银娟钱柳青
Owner GUANGZHOU FULENGEN
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