Application of miRNA-324-5p and its activators in the preparation of clinical markers and therapeutic drugs for inflammatory bowel disease and inflammatory bowel disease-related colorectal cancer

A technology of inflammatory bowel disease and activators, applied in the field of biomedicine, to achieve the effect of alleviating intestinal inflammation

Active Publication Date: 2020-06-26
李建明 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Purpose of the invention: The technical problem to be solved by the present invention is to overcome the deficiency of drugs for treating inflammatory bowel disease and inflammatory bowel disease-related colorectal cancer in the prior art and the limitations of clinical diagnostic markers for inflammatory bowel disease. In-depth research on its downstream miRNA, on the basis of the original research on miRNA-324-5p, through a large number of experimental screening, a new use of miRNA-324-5p was developed

Method used

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  • Application of miRNA-324-5p and its activators in the preparation of clinical markers and therapeutic drugs for inflammatory bowel disease and inflammatory bowel disease-related colorectal cancer
  • Application of miRNA-324-5p and its activators in the preparation of clinical markers and therapeutic drugs for inflammatory bowel disease and inflammatory bowel disease-related colorectal cancer
  • Application of miRNA-324-5p and its activators in the preparation of clinical markers and therapeutic drugs for inflammatory bowel disease and inflammatory bowel disease-related colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Experiment of miR-324-5p as a diagnostic marker for inflammatory bowel disease

[0028] 1. Culture of colorectal tumor cells in vitro:

[0029] Three wild-type colorectal cancer cells (RKO, DLD1, HCT116) and their corresponding DICER gene knockout cells (DICER - / -) cultured in 1640 medium containing 10% FBS, placed in a 37°C, 5% CO2 incubator to grow, to the logarithmic growth phase for the test; when the cells grow to a degree of confluence greater than 80%, subculture: PBS buffer Wash 2 times with trypsin, the volume of trypsin covers the cell surface, incubate at 37°C for 1 min, and observe under the microscope under bright field. When the cells shrink and the edge is clear, discard the digestion solution, add complete medium to stop digestion, and blow with a pipette The cells on the wall of the flask were made into a single cell suspension, which was inoculated into a new petri dish for culture.

[0030] 2. Breeding and observation of mice

[0031] The...

Embodiment 2

[0039] Example 2 Animal experiment of miRNA-324-5p activator in treating inflammatory bowel disease

[0040] 1. Experimental grouping:

[0041] 6-8 weeks old, male wild-type mice (Dicer loxp / + ) and Dicer heterozygous knockout mice (Dicer loxp / + &Villin Cre ) were randomly divided into three groups: control drinking group (water), DSS induction group (DSS), DSS induction and miR-324-5p activator intraperitoneal injection group (DSS+AgomiR-324-5p).

[0042] 2. Experimental method:

[0043] Dissolve DSS (MW: 36000-50000) in drinking water to prepare a 3% DSS solution. Littermate wild-type mice (Dicer loxp / + ) and Dicer heterozygous knockout mice (Dicer loxp / + &Villin Cre ) mice were free to drink distilled water and distilled water containing 3% DSS respectively. On the second day, AgomiR-324-5p with a concentration of 10nM or its control was injected intraperitoneally at 200 μl / mouse, and the daily status of mice such as eating, drinking and activities were observed eve...

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Abstract

The invention discloses application of miRNA-324-5p and an activator thereof in the preparation of a clinic marker for inflammatory bowel disease and colorectal cancer relevant with the inflammatory bowel disease as well as a drug for treating the inflammatory bowel disease and the colorectal cancer relevant with the inflammatory bowel disease. The screening of a large number of experiments finds that the severe inflammatory bowel disease of mice is spontaneously caused along with the knockout of intestinal mucosa epithelial cell DICER genes; meanwhile, miRNA-324-5p as one, with the most remarkable change, of miRNAs can be used as a clinical test marker of the inflammatory bowel disease. More particularly, by intraperitoneally injecting a miRNA-324-5p activator into a DSS-induced mouse colonitis model and a human inflammatory bowel disease simulated model, the symptom of mouse acute colonitis can be remarkably relieved, so that the use of the miRNA-324-5p activator in the treatment of patients with the inflammatory bowel disease and the colorectal cancer relevant with the inflammatory bowel disease is indicated.

Description

technical field [0001] The invention relates to a new application of microRNA molecules, in particular to a new application of miRNA-324-5p, and belongs to the technical field of biomedicine. Background technique [0002] Inflammatory bowel disease (IBD) is a group of chronic nonspecific intestinal inflammation of unknown etiology, including ulcerative colitis (UC) and Crohn's disease (CD). With the deepening of research, people gradually realize that IBD is a risk factor for colorectal cancer, and it will make colorectal cancer younger. In the past five years, the number of IBD patients in my country has been eight times that of the same period in the 1990s, and it is urgent to receive widespread attention. MicroRNA (microRNA) is a class of single-stranded RNA that regulates gene expression at the post-transcriptional level, with a length of about 22bp. miRNA regulates gene expression by mediating mRNA degradation and inhibiting mRNA transcription. Studies have shown that...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K31/7088A61P1/00
CPCA61K31/7088A61K45/00
Inventor 李建明孙丽娜支政邢澄
Owner 李建明
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