Biomarker for lung adenocarcinoma diagnosis and treatment

A lung adenocarcinoma and drug technology, applied in the field of biomedicine, can solve problems such as poor prognosis

Inactive Publication Date: 2017-05-24
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these have not fundamentally changed the current situation of high mortality from lung cancer, especially the overall prognosis of lung cancer patients lacking targeted genes is still poor, and the 5-year survival rate is still hovering around 15%.

Method used

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  • Biomarker for lung adenocarcinoma diagnosis and treatment
  • Biomarker for lung adenocarcinoma diagnosis and treatment
  • Biomarker for lung adenocarcinoma diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Screening for Gene Markers Related to Lung Adenocarcinoma

[0070] 1. Sample collection

[0071] 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.

[0072] 2. RNA sample preparation (using miRNA kit for operation)

[0073] Introduce liquid nitrogen into the mortar, take the tissue obtained above and put it into the mortar, cut it into pieces in liquid nitrogen ...

Embodiment 2

[0095] Example 2 QPCR sequencing to verify the differential expression of the FIBIN gene

[0096] 1. Large-sample QPCR verification of differential expression of FIBIN 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.

[0097] 2. The RNA extraction steps are as described in Example 1.

[0098] 3. Reverse transcription

[0099] 1) Reaction system:

[0100] 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.

[0101] Continue to add the following components to the 12 μl reaction solution:

[0102] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and centrifuge;

[0103] 2) Reverse transcription reaction conditions

[0104] 5 minutes at 25°C, 60 minutes at 42°C, and 5 min...

Embodiment 3

[0121] Example 3 FIBIN overexpression

[0122] 1. Cell culture

[0123] 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.

[0124] 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%.

[0125] 2. Construction of gene overexpression vector

[0126] Synthesize specific PCR amplification primers according to the cDNA sequence of FIBIN (as shown in SEQ ID NO.1), and the primer sequences are as follows:

[0127] Forward primer: 5'-CCGAAGCTTGCCACCATGGTGTTTCTGAAGTTC-3' (SEQ ID NO.7)

[0...

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Abstract

The invention discloses a biomarker for lung adenocarcinoma diagnosis and treatment. The expression of marker FIBIN in a lung adenocarcinoma tissue is down-regulated, and the early diagnosis of the lung adenocarcinoma can be performed by detecting the expression level of the FIBIN. The invention also proves that the up-regulation of the expression of the FIBIN can effectively inhibit the proliferation and invasion of a lung adenocarcinoma cell through the experiment, thereby prompting that the FIBIN can be used for treating the lung adenocarcinoma or the lung adenocarcinoma metastasis. The invention further discloses a method for screening a potential substance for treating the lung adenocarcinoma, namely, the potential substance can up-regulate or increase the expression level or the expression product activity of the FIBIN.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a biomarker for diagnosis and treatment of lung adenocarcinoma, in particular to FIBIN as the biomarker. Background technique [0002] Lung cancer is one of the tumors with the highest morbidity and mortality worldwide. Its incidence is still at its peak in many countries. According to the latest WHO statistics, 1.59 million patients worldwide die of lung cancer every year. According to data released by my country Cancer Registry Center in 2016: In 2015, there were about 733,000 new cases of lung cancer in my country (509,300 males and 224,000 females), and the number of lung cancer deaths in my country was 610,200 (432,400 males and 177,800 females) in the same period. . Lung cancer has become the tumor with the highest morbidity and mortality in my country, especially in recent years, the prevalence of lung cancer in women has increased significantly. At present, surgery, chemotherap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00C12Q2600/106C12Q2600/158G01N33/57423G01N33/68G01N2333/75
Inventor 杨承刚宋宏涛
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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