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
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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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