miRNA associated with colorectal cancer and application thereof

A technology for colorectal cancer and its application, which is applied in the field of miRNA and its application, can solve the problems of easy to miss micro-metastasis, time-consuming, and inaccurate tumor staging, etc., and achieve the goal of improving healing power, reducing patient mortality, and speeding up diagnosis and treatment Effect

Inactive Publication Date: 2016-06-01
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 colorectal cancer

[0025] (1) Establish a standardized and normalized colorectal 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. Tissues and plasma are stored at -80°C, and the clinical data of the collected samples (such as: hospitalization number, pathology number, tissue location, tumor stage and grade, operation method, chemotherapy regimen, follow-up) and experimental data (such as: extraction miRNA concentration, spectrophotometry, electrophoresis pictures, quantitative PCR experiment results) were counted and organized, and a colorectal cancer miRNA database available for sharing wa...

Embodiment 2

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

[0040]Fluorescent quantitative PCR was used to detect the expression of miR-1290 in the plasma of 219 colorectal cancer patients and 45 age- and gender-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 BestarqPCRRTKit instructions, synthesize cDNA, and store it at -20°C after co...

Embodiment 3

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

[0050] This embodiment provides a kit for differentiating colorectal cancer samples from normal samples in a subject's blood sample. The kit includes: a detection reagent suitable for detecting miR-1290; A device for detecting the amount of miR-1290 detected by the detection reagent.

[0051] The kit is a real-time fluorescence quantitative PCR detection kit for colorectal cancer, which includes a test strip with an oligonucleotide probe spotted on it, and the sequence of the oligonucleotide probe 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 PCR d...

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Abstract

The invention belongs to the field of nucleic acid diagnosis and treatment and specifically relates to miRNA associated with colorectal cancer and an application thereof. The miRNA associated with colorectal cancer is miR-1290. The invention provides an application of the miR-1290 used in preparing a kit for distinguishing a colorectal cancer sample and a normal sample in a blood sample of a subject. The invention confirms for the first time that the miR-1290 is a colorectal cancer-associated miRNA diagnosis marker, and a novel colorectal cancer progression detection index is established. On this basis, the invention provides a new application of the miR-1290 in initial diagnosis and rapid detection of colorectal cancer and a related universally 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 colorectal cancer and its application. Background technique [0002] Colorectal cancer is one of the most common malignant tumors that seriously endanger people's health. It ranks first in cancer incidence and second in mortality in developed countries, and fourth in cancer mortality in my country, and its incidence is increasing year by year. In Guangzhou, the crude incidence and mortality rates of colorectal 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 colorectal cancer. The phase drops to 75±%, the C phase is less than 50%, and the D phase is less than 10%. B, C, and D three-stage cases accounted for 96.17%, indic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李华平张金阔吴银娟钱柳青
Owner GUANGZHOU FULENGEN
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