Application of LINC02085 gene and detection method of LINC02085 gene

A gene and gene expression technology, applied in genetic engineering, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of RA lack of diagnostic markers, lack of targeting and pertinence of therapeutic drugs, etc.

Active Publication Date: 2021-02-12
FIRST AFFILIATED HOSPITAL OF ANHUI UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of the lack of diagnostic markers for RA and the lack of targeting and pertinence of therapeutic drugs, the present invention provides an application of LINC02085 gene and its detection method

Method used

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  • Application of LINC02085 gene and detection method of LINC02085 gene
  • Application of LINC02085 gene and detection method of LINC02085 gene
  • Application of LINC02085 gene and detection method of LINC02085 gene

Examples

Experimental program
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Effect test

Embodiment 1

[0021] In order to explore the expression of LINC02085 in PBMCs of RA patients.

[0022] After the project has passed the review of the Ethics Committee, the applicant will systematically study the new diagnostic and therapeutic targets of human RA, extract PBMCs from the peripheral blood of RA patients, and detect the expression of the LINC02085 gene by RT-qPCR.

[0023] In this embodiment, a total of 30 blood samples from RA patients and 30 blood samples from healthy people in the control group were collected, a total of 60 samples (see Table 1 for specific information from 30 patients, and see Table 2 for sample characteristics)

[0024] Table 1 Sample information

[0025]

[0026]

[0027] Table 2 Sample Characteristics

[0028]

[0029] In this embodiment, 30 cases of RA and 30 cases of healthy control group PBMCs were detected by RT-qPCR technology. The specific detection steps are as follows:

[0030] 1. Extraction of PBMCs from RA patients

[0031] At room ...

Embodiment 2

[0048] Example 2 LINC02085 overexpression sequence and small interfering sequence and application thereof

[0049] This example designs the full-length sequence of LINC02085, and synthesizes its specific overexpression sequence pcDNA3.1-LINC02085 and small interfering sequence si-LINC02085 (its nucleotide sequence is shown in SEQ ID NO.3), and infects FLS, Make LINC02085 gene overexpression and small interference in cells, and further apply the overexpression and small interference gene carrier to regulate the inflammatory response of FLS.

[0050] The specific virus packaging and cell infection methods are as follows:

[0051] (1) When 293T cells are cultured in a 6cm dish until 80-90% confluent, discard the culture medium and wash the cells twice with 3mL PBS; (2) Add 1mL Trypsin-EDTA solution, mix well, and carefully suck off the trypsin solution , placed at 37°C for 3 minutes; (3) Add 2 mL of DMEM culture medium containing 10% FBS, and pipette to make the cells form a sin...

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Abstract

The invention discloses application and a detection method of an LINC02085 gene, and belongs to the technical field of molecular biology of rheumatism. According to the invention, a specific PCR primer is designed and synthesized according to the extracted total RNA sequence, and an Rt-qPCR technology is utilized to find that the expression of LINC02085 in peripheral blood mononuclear cells of a rheumatoid arthritis (RA) patient is significantly increased, which proves that the LINC02085 can be applied to the preparation of a preparation for auxiliary diagnosis, treatment or prognosis evaluation of rheumatoid arthritis. The pcDNA3.1-LINC02085 sequence for specific overexpression of the gene and the si-LINC02085 sequence for small interference of the gene are designed and synthesized at thesame time, proliferation of synovial fibroblasts (FLS) and inflammatory response of FLS induced by TNF-alpha can be remarkably inhibited by regulating and controlling a PI3K/AKT signal channel, and the gene can become a molecular targeted therapy tool for RA.

Description

technical field [0001] The invention belongs to the technical field of molecular biology of rheumatic diseases, and relates to the expression, application and detection field of a long-chain non-coding RNA (LINC02085), in particular to an application of the LINC02085 gene and a detection method thereof. Background technique [0002] Rheumatoid arthritis (RA) is characterized by symmetry, peripherality, and polyarthritis, which is chronic, progressive, and aggressive. It is an autoimmune, inflammatory disease that can cause systemic multisystem damage disease. The global incidence of RA is high, and the incidence of women is higher than that of men. The formation of synovitis and pannus is the basic pathological change of RA. A large number of previous studies have concluded that the pathogenesis of RA is complex, and its occurrence and development may be closely related to heredity, surrounding environment, immune dysfunction, and platelet activation. If RA patients are no...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11C12N15/113
CPCC12N15/113C12N2310/141C12Q1/6883C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 刘健姜辉万磊文建庭孙艳秋
Owner FIRST AFFILIATED HOSPITAL OF ANHUI UNIV OF CHINESE MEDICINE
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