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Application of lncRNA (long no-coding RNA) in diagnosis and tretament of lung adenocarcinoma

A lung adenocarcinoma and reagent technology, applied in the field of biomedicine, can solve the problem of lncRNA not being discovered

Active Publication Date: 2019-03-15
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although lncRNAs play an important role in the development of lung adenocarcinoma, and some lncRNAs can be used as indicators for the diagnosis and prognosis of lung adenocarcinoma, there are still a large number of lncRNAs that have not been discovered, and further research on the expression and regulation of lncRNAs in lung adenocarcinoma is still needed Mechanisms in Tumor Development

Method used

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  • Application of lncRNA (long no-coding RNA) in diagnosis and tretament of lung adenocarcinoma
  • Application of lncRNA (long no-coding RNA) in diagnosis and tretament of lung adenocarcinoma
  • Application of lncRNA (long no-coding RNA) in diagnosis and tretament of lung adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1 Screening for Gene Markers Related to Lung Adenocarcinoma

[0091] 1. Sample collection

[0092] Surgical specimens of 4 lung adenocarcinoma tissues and paired paracancerous tissues (≥5cm from the tumor edge) were collected respectively. All patients had not received chemotherapy, radiotherapy, targeted therapy, tumor immunotherapy and other treatments before surgery, and no other For neoplastic diseases, autoimmune diseases and severe chronic diseases, all patients have given informed consent and approved by the organizational ethics committee.

[0093] 2. Preparation of RNA samples

[0094] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific operation was carried out according to the instructions.

[0095] 3. Total RNA quantification and purity analysis

[0096] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity o...

Embodiment 2

[0114] Example 2 QPCR sequencing to verify the differential expression of LINC01983 gene

[0115] 1. Large-sample QPCR verification of differential expression of LINC01983 gene. According to the sample collection method in Example 1, 35 cases of lung adenocarcinoma paracancerous tissues and 35 cases of lung adenocarcinoma tissues were selected.

[0116] 2. RNA extraction

[0117] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0118] 3. QPCR

[0119] 1) Reverse transcription reaction

[0120] Use FastQμant cDNA First Strand Synthesis Kit (Product No.: KR106) for lncRNA reverse transcription, first remove the genomic DNA reaction, add 5×gDNA Bμffer 2.0μl, total RNA 1μg, add RNase Free ddH 2 O to bring the total volume to 10 μl, and heat in a water bath at 42°C for 3 min.

[0121] 10× Fast RT Bμffer 2.0μl, RT Enzyme Mix 1.0μl, FQ-RT Primer Mix 2.0μl, RNase Free ddH 2 O 5.0 μl, after mixing, add t...

Embodiment 3

[0141] Example 3 Silencing of the LINC01983 gene

[0142] 1. Cell culture

[0143] Human lung adenocarcinoma cell line A549 was incubated in RPMI1640 medium containing 10% fetal bovine serum and 1% P / S at 37°C, 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0144] 2. Design of siRNA

[0145] Design siRNA for the sequence of LINC01983 gene, the designed siRNA sequence is as follows:

[0146] Sequence of negative control siRNA-NC:

[0147] Sense strand: 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.5),

[0148] Antisense strand: 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.6);

[0149] siRNA1:

[0150] Sense strand: 5'-ACUCAAAUUGGAAAGCUUGAC-3' (SEQ ID NO.7),

[0151] Antisense strand: 5'-CAAGCUUUCCAAUUUGAGUCU-3' (SEQ ID NO.8);

[0152] siRNA2:

[0153] Sense strand: 5'-UUUUUUGCUUCAUUUGCAGGU-3' (SEQ ID NO.9),

[0154] Antisense strand: 5'-CUGCAAAUGAAG...

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Abstract

The invention discloses an application of lncRNA (long no-coding RNA) which is LINC01983 in diagnosis and tretament of lung adenocarcinoma. Significant up-regulation of expression of the LINC01983 inlung adenocarcinoma is found for the first time, and large-sample QPCR experiments further prove that the LINC01983 can be applied to clinical diagnosis of the lung adenocarcinoma as a potential molecular target. The invention also discloses the LINC01983 related to proliferation, migration and invasion of lung adenocarcinoma cells, which indicates that the LINC01983 can be applied to clinical diagnosis of the lung adenocarcinoma as the potential molecular target.

Description

technical field [0001] The invention belongs to the field of biomedicine, relates to the application of lncRNA in the diagnosis and treatment of lung adenocarcinoma, and specifically relates to the application of LINC01983 in the diagnosis and treatment of lung adenocarcinoma. Background technique [0002] Lung cancer is a global problem, devouring the lives of millions of people every year, and is the main cause of cancer death worldwide (Denisenko TV, Budkeveh IN, Zhivotovsky B. Cell death-based treatment of lung adenocarcinoma[J]. Cell Death Dis.2018,9(2):117.). Although the incidence of lung cancer is the second among male and female malignant tumors, the mortality rate ranks first among all malignant tumors (RebeccaL. Siegel, MPH, et al. Cancer statistics, 2018. CA Cancer J C1in. 2018 ; 68:7-30.). In recent years, with the development of the economy, the improvement of sanitary conditions, and the continuous improvement of medical diagnosis and treatment, the diagnosi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11A61K45/00A61P35/00A61P35/04
CPCA61K45/00A61P35/00A61P35/04C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 杨承刚高舒欣魏琳
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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