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Application of miRNA in liver cirrhosis diagnosis and treatment

A 1. miRNA-548a-3p, patient technology, applied in the field of biomedicine, can solve problems such as inducing cancer and abnormal gene expression

Active Publication Date: 2015-12-23
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The abnormal expression of miRNA directly leads to the abnormal expression of some genes related to the occurrence of diseases, and induces the occurrence of cancer.

Method used

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  • Application of miRNA in liver cirrhosis diagnosis and treatment
  • Application of miRNA in liver cirrhosis diagnosis and treatment
  • Application of miRNA in liver cirrhosis diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Screening of miRNAs associated with liver cirrhosis

[0045] 1. Sample acquisition: 10 biopsy samples from healthy people and 10 biopsy samples from patients with liver cirrhosis were collected. The acquisition of all the above samples was approved by the ethics committee of the organization.

[0046] 2. Extraction of total RNA from samples

[0047] Total RNA in advance using QIAGEN Tissue RNA Extraction Kit. Specific steps are as follows:

[0048] 1) In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh about 20 mg of the biopsy tissue sample, and grind it to powder with a pestle;

[0049] 2) Transfer the sample to a 2ml centrifuge tube without RNase;

[0050] 3) Add 300 μl Lysissolution, place in a homogenizer, and grind thoroughly for 1-5 minutes;

[0051] 4) 12000g, 4°C, centrifuge for 10min, transfer the supernatant to a new 1.5ml centrifuge tube;

[0052] 5) Add 600 μl RNase-FreeWate...

Embodiment 2

[0075] Example 2 QPCR verification of differentially expressed miRNA-548a-3p

[0076] 1. Select miRNA-548a-3p according to the detection results of the miRNA chip for large sample QPCR verification. According to the sample collection method in Example 1, 80 cases of biopsy tissue samples from patients with liver cirrhosis and 80 biopsy samples from healthy people were selected.

[0077] 2. The RNA extraction process is the same as in Example 1.

[0078] 3. Reverse transcription:

[0079] 1) Prepare the reaction system according to Table 1

[0080] Table 1 reaction system

[0081] RNA template

1μg

10× buffer

2μl

dATP (10mM)

2μl

polyA polymerase

0.5μl

RNase inhibitor

0.5μl

wxya 2 o

Make up to 20μl

[0082] Incubate at 37°C for 1h.

[0083] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0084] 3) Immediately incubate on ice for 2 ...

Embodiment 3

[0101] Example 3 Study on the effect of miRNA-548a-3p on the proliferation ability of liver cirrhosis cells

[0102] 1. Design and synthesis of mimics targeting miRNA-548a-3p

[0103] According to the sequence information of miRNA-548a-3p, miRNA-548a-3p mimics and random control sequences were designed and synthesized by Dalian Bao Biotechnology Co., Ltd.

[0104] 2. Cell culture

[0105] Hepatic stellate cells LX-2 were incubated with DMEM high-glucose medium containing 10% fetal bovine serum at 37°C, 5% CO 2 cultured in an incubator.

[0106] 3. Cell transfection

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Abstract

The invention discloses application of miRNA in liver cirrhosis diagnosis and treatment. miRNA is miRNA-548a-3p. By detecting the expression level of miRNA-548a-3p, miRNA-548a-3p can be used for diagnosing whether a patient has liver cirrhosis or is in the liver cirrhosis risk. As is shown in tests, the expression level of miRNA-548a-3p in a patient with liver cirrhosis is obviously lower than that of miRNA-548a-3p in a healthy person. On this basis, miRNA-548a-3p can also be used for preparing liver cirrhosis treatment medicine. Through miRNA-548a-3p, the sensitivity and specificity of liver cirrhosis diagnosis are greatly improved, and meanwhile a new target spot is provided for gene treatment of liver cirrhosis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of miRNA in the diagnosis and treatment of liver cirrhosis, in particular to the application of miRNA-548a-3p in the diagnosis and treatment of liver cirrhosis. Background technique [0002] Liver cirrhosis is a common clinical chronic progressive liver disease, which is diffuse liver damage caused by long-term or repeated effects of one or more causes. In my country, most of them are post-hepatitis cirrhosis, and a small part are alcoholic cirrhosis and schistosome cirrhosis. Liver cirrhosis is the pathological process of abnormal hyperplasia of fibrous connective tissue in the liver when liver cells undergo necrosis and inflammatory stimulation. Excessive fibrosis causes liver atrophy and hardening, which eventually leads to decompensation of liver cirrhosis or liver failure, resulting in death of the patient. In the early stage, there are no obvious symptoms due to the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K45/00A61P1/16
CPCA61K45/00C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 高倩倩杨承刚
Owner BEIJING MEDINTELL BIOMED CO LTD
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