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Application of biomarker as target to diagnosis and treatment of lung adenocarcinoma

A kind of lung adenocarcinoma, application technology, applied in the field of biomedicine

Active Publication Date: 2017-05-31
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Available data also suggest that biomarkers may be easier to discover and may benefit from in lung adenocarcinoma

Method used

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  • Application of biomarker as target to diagnosis and treatment of lung adenocarcinoma
  • Application of biomarker as target to diagnosis and treatment of lung adenocarcinoma
  • Application of biomarker as target to diagnosis and treatment of lung adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1 Screening for Gene Markers Related to Lung Adenocarcinoma

[0080] 1. Sample collection

[0081] Paracancerous tissue and lung adenocarcinoma tissue samples were collected from 8 cases of lung adenocarcinoma. The tumor tissue specimens of lung adenocarcinoma were collected from the main tumor area, which was located at the junction of the middle and outer 1 / 3 of the tumor mass and normal tissue, and the obvious necrosis and calcification in the tumor center and normal lung tissue around the tumor were excluded; the normal lung tissue specimens adjacent to cancer were obtained from tumor There is no obvious change in the part above 5cm from the edge. All the above specimens were obtained with the consent of the organizational ethics committee.

[0082] 2. RNA sample preparation (using E.Z.N.A. miRNA kit for operation)

[0083] Introduce liquid nitrogen into the mortar, take the tissue obtained above and put it into the mortar, cut it into pieces in liquid ...

Embodiment 2

[0105] Example 2 QPCR sequencing to verify the differential expression of the CLINT1 gene

[0106] 1. Large-sample QPCR verification of differential expression of CLINT1 gene. According to the sample collection method in Example 1, 50 cases of lung adenocarcinoma paracancerous tissues and 50 cases of lung adenocarcinoma tissues were selected.

[0107] 2. The RNA extraction steps are as described in Example 1.

[0108] 3. Reverse transcription

[0109] 1) Reaction system:

[0110] RNA template 1 μl, random primer 1 μl, double distilled water was added to 12 μl, mixed, centrifuged at low speed, 65 ° C for 5 min, and then cooled on ice.

[0111] Continue to add the following components to the 12 μl reaction solution:

[0112] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and centrifuge;

[0113] 2) Reverse transcription reaction conditions

[0114] 5 minutes at 25°C, 60 minutes at 42°C, and 5 m...

Embodiment 3

[0131] Silence of embodiment 3CLINT1 gene

[0132] 1. Cell culture

[0133] 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.

[0134] The cells in the culture flask were digested with trypsin and seeded in a 6-well plate to ensure that the number of cells was 2-8×10 5 cells / well, add cell culture medium. Overnight, observe the cell density the next day, and transfection can be performed when the cell density is above 70%.

[0135] 2. Design of siRNA

[0136] Negative control siRNA sequence (siRNA-NC):

[0137] The sense strand is 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.7)

[0138] The antisense strand is 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.8)

[0139] siRNA-1:

[0140] The sense strand is 5'-UUAUCACUGAAUGGAAAAGCA-...

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Abstract

The invention discloses application of a biomarker as a target to the diagnosis and the treatment of lung adenocarcinoma. Particularly, the biomarker is CLINT1. The worldwide incidence rate of the lung adenocarcinoma continuously increases; an effective biomarker is needed to diagnose and / or treat the lung adenocarcinoma. The application is proven through an experiment; the expression, in a patient suffered from the lung adenocarcinoma, of the CLINT1 is up-regulated; the multiplication and the invasion of a lung adenocarcinoma cell can be inhibited by decreasing the expression of a CLINT1 gene. Prompted, the CLINT1 disclosed by the invention can be used as a marker for the early diagnosis of the lung adenocarcinoma; meanwhile, the gene can be also used as a potential drug target spot for treating the lung adenocarcinoma; the foundation is provided for the accurate medical treatment of the lung adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of a biomarker as a target in the diagnosis and treatment of lung adenocarcinoma, and the specific biomarker is CLINT1. Background technique [0002] Lung cancer is one of the tumors with the highest morbidity and mortality worldwide, seriously endangering human health. Lung cancer can be divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Among them, NSCLC accounts for about 80%-85% of the total number of lung cancers, and its disease develops rapidly, with rapid metastasis and high recurrence rate. Most of the patients have developed to the middle and late stages when they are discovered, the prognosis is poor, and the 5-year survival rate is less than 15%. The recurrence rate of NSCLC patients who received early treatment was as high as 40%, and more than 70% of the patients were found to have local invasion or distant metastasis and co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/713A61K45/00A61P35/00C12Q1/68G01N33/68G01N33/577
CPCA61K31/713A61K45/00A61K48/005C12Q1/6886C12Q2600/118G01N33/577G01N33/6884G01N2800/12
Inventor 边洋李曙光
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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