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Application of circular RNA as osteoarthritis marker

An osteoarthritis and annular technology, applied in the field of biomedicine, can solve problems such as cartilage degeneration of regenerated cartilage tissue, and achieve the effect of significant significance and good application prospects.

Active Publication Date: 2021-08-13
GUANGDONG HOSPITAL OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main prevention and repair measures are: mesenchymal stem cell transplantation to promote cartilage differentiation, autologous cartilage or biomaterial transplantation to fill the cartilage defect site, although a certain curative effect has been achieved, but there are still regenerated cartilage tissue and cartilage degeneration at the edge of the defect And other issues

Method used

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  • Application of circular RNA as osteoarthritis marker
  • Application of circular RNA as osteoarthritis marker
  • Application of circular RNA as osteoarthritis marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] This example studies the expression of circular RNAhsa_circ_0000423 in normal volunteers and patients with osteoarthritis.

[0047] Using Array Human Circular RNA gene chip sequencing to detect 4 cases of cartilage tissue samples from normal (Normal) and osteoarthritis (OA) patients, and conducted bioinformatics analysis, the expression profiles of circRNAs that were found to be different in the samples were found. The results are as follows: figure 1 shown. in, figure 1 Figure A in the middle is the clustering diagram of the expression profile of circRNAs, which shows that there are significant differences in the expression profiles of circRNAs between the two groups, and the expression profiles in each group are relatively consistent; figure 1 Figure B in the middle is a scatter plot of circRNAs, and the lines outside the lines are circRNAs with expression difference > 2; figure 1 Middle C is the volcano map of differentially expressed circRNAs, and the dark dots sh...

Embodiment 2

[0049] 1. Biological characteristics of circular RNAhsa_circ_0000423

[0050] Circular RNA hsa_circ_0000423 is used as an OA marker, including 1138bp, and its full nucleic acid sequence is shown in SEQ INNO.1.

[0051] 2. Circular verification of circular RNAhsa_circ_0000423

[0052] The specific primer of the OA marker based on circular RNAhsa_circ_0000423: including the forward primer of hsa_circ_0000423 and the reverse primer of hsa_circ_0000423, the nucleotide sequence of the forward primer of hsa_circ_0000423 is shown in SEQ IN NO: 2, the reverse of hsa_circ_0000423 The nucleotide sequence of the primer is shown in SEQ INNO:3.

[0053] RNase R enzyme digestion: use GENESEED RNase R digestive enzyme, configure 10 μL of reaction system according to 2 μg total RNA + 0.5 μL RNase R + 1 μL Reaction Buffer, and use RNase free H for insufficient volume 2 O supplementation, 37°C, incubation for 20 minutes, reverse transcription at 70°C for 10 minutes to inactivate the enzyme, a...

Embodiment 3

[0063] The effect of hsa_circ_0000423 on the biological function of OA was verified by transfecting the overexpressed hsa_circ_0000423 plasmid, which was provided by Shanghai Heyuan Biotechnology Co., Ltd.

[0064] By transfecting the overexpressed hsa_circ_0000423 plasmid into human chondrocytes, Western Blot technology was used to detect the protein levels of related metabolic products (MMP13 and COL-2) in human chondrocytes. The specific steps are as follows:

[0065] 1. Transfection

[0066] 1. One hour before transfection, suck off the medium in the 6-well plate, replace with 2mL opti-MEM medium per well or the serum-free medium corresponding to the cells, and place at 37°C, 5% CO 2 in a constant temperature incubator.

[0067] 2. According to the number of transfected plasmids, prepare enough EP tubes, and clearly mark the name of the plasmid on the tube cap. One of the EP tubes is the main tube of opti-MEM medium, and one EP tube is the PEI transfection reagent (or Tra...

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Abstract

The invention belongs to the technical field of biomedicine and particularly relates to application of circular RNA as an osteoarthritis marker. The circular RNA is has_circ_0000423 and has a nucleotide sequence represented by SEQ ID NO: 1. According to the application, researches discover that compared with normal cartilage tissue, hsa_circ_0000423 is of high expression in cartilage tissue of an osteoarthritis sufferer, so that judgment on whether a testee has the high risk of suffering from osteoarthritis or not is contributed through detecting the content of the hsa_circ_0000423, and the significance in clinical auxiliary diagnosis and evaluation on the osteoarthritis is remarkable. Simultaneously, through further researching biological functions of cartilage cells, diagnosis and treatment values of the hsa_circ_0000423 during occurrence and development of OA are discovered, so that a critical theoretical basis and intervention target is provided for diagnosis and treatment on the disease, and thus, the circular RNA has an important potential value and a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of circular RNA has_circ_0000423 as a marker of osteoarthritis. Background technique [0002] Osteoarthritis (OA) is an imbalance in the normal synthesis and degradation of articular chondrocytes, extracellular matrix, and subchondral bone under the action of mechanical and biological factors, resulting in articular cartilage degeneration, fibrosis, and subchondral bone sclerosis. , inflammatory changes and osteophyte formation, joint pain, limited mobility, deformity and other clinical manifestations of a disease. Osteoarthritis is a long-standing orthopedic clinical problem that needs to be solved urgently. It often occurs in the elderly, and often involves the spine, hip joints, knee joints and other joints with heavy loads in the bone joints of the whole body, resulting in stiffness and deformation of the affected joints. Joint function is ser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11A61K45/00A61P19/02
CPCC12Q1/6883A61K45/00A61P19/02C12Q2600/178C12Q2600/136C12Q2600/118C12Q2600/158
Inventor 李兴林定坤侯宇陈树东侯永辉罗丹詹吉恒栾继耀
Owner GUANGDONG HOSPITAL OF TRADITIONAL CHINESE MEDICINE
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