Marker for identifying benign and malignant pulmonary nodules by using exosome protein and application of marker

A technology of pulmonary nodules and exosomes, which is applied in the field of exosomal proteins as markers for distinguishing benign and malignant pulmonary nodules, and can solve the problems of unreported FGB expression in benign and malignant pulmonary nodules, etc.

Inactive Publication Date: 2019-03-15
HUADONG HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current study has not yet reported the expression of FGB in plasma exosomes of patients with pulmonary nodules and the role of FGB in differentiating benign and malignant pulmonary nodules.

Method used

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  • Marker for identifying benign and malignant pulmonary nodules by using exosome protein and application of marker
  • Marker for identifying benign and malignant pulmonary nodules by using exosome protein and application of marker
  • Marker for identifying benign and malignant pulmonary nodules by using exosome protein and application of marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] see Figure 1-3 , the present invention provides the following technical scheme: an exosomal protein as a marker for differentiating benign and malignant pulmonary nodules and its application, comprising the following steps:

[0027] S1. Search for proteins differentially expressed in plasma exosomes of patients with benign and malignant pulmonary nodules;

[0028] S2, screening differential proteins;

[0029] S3. To verify the role of FGB in the differential diagnosis of benign and malignant pulmonary nodules;

[0030] S4. Analysis of relative expression

[0031] In the present invention, non-invasive liquid biopsy methods are used to find molecular markers for distinguishing benign and malignant pulmonary nodules, and through mass spectrometry detection and subsequent bioinformatics analysis, it is shown that the molecular marker FGB of the present invention is the diagnosis and clinical efficacy of pulmonary nodules The observation provides a new molecular target....

Embodiment 2

[0054] Example 2: Screening of target markers of the present invention

[0055] In order to screen the differences of exosomal proteins in patients with benign and malignant pulmonary nodules, a protein profile study was carried out to screen out a series of differential proteins ( figure 2 ).

[0056] The experimental steps are as follows:

[0057] (1) Cleavage and quantification of protein:

[0058] Label the collected exosomes, add 100 μL of SDT lysate, boil water bath for 10 min, and sonicate (100 W, work for 10 s, pause for 10 s, cycle 10 times). Centrifuge at 13400rpm for 30min, and take the supernatant. BCA method for protein quantification.

[0059] (2) SDS-PAGE experiment of protein protein

[0060] Take 20 μg of protein samples from each group and add 5X loading buffer at 5:1 (v / v), put in boiling water bath for 5 minutes, centrifuge at 14000 g for 10 minutes to get the supernatant, and perform 12.5% ​​SDS-PAGE electrophoresis. Conditions: constant current 15m...

Embodiment 3

[0081] Example 3: Verification of screening markers

[0082] In the stage of screening differential proteins, plasma exosome samples from 30 patients with malignant pulmonary nodules and 10 patients with benign pulmonary nodules were included.

[0083] The inclusion criteria for patients are:

[0084] 1. Patients diagnosed with pulmonary nodules by imaging from April to July 2016;

[0085] 2. The patient agrees to receive surgical treatment and resect the lesion;

[0086] Exclusion criteria:

[0087] 1. Exclude patients with other tissue system tumors;

[0088] 2. Exclude other lung disease patients with non-substantial pulmonary lesions.

[0089] Blood samples were collected from patients who met the above criteria, and 5-8mL of peripheral blood was drawn, placed in an EDTA anticoagulant tube, centrifuged twice, and the supernatant was taken for exosome extraction.

[0090] The inventors analyzed the protein profiles of the two groups of samples, and screened out a serie...

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Abstract

The invention belongs to the technical field of cancer molecular diagnosis technique, in particular to a marker for identifying benign and malignant pulmonary nodules by using an exosome protein and an application of the marker. The marker comprises the following steps: S1, looking for differentially expressed proteins of exosomes in plasmas of patients suffering from benign and malignant pulmonary nodules; S2, screening the differential proteins; S3, verifying the differential diagnosis effect of FGB on the benign and malignant pulmonary nodules; and S4, analyzing the relative transcript level. The non-invasive liquid-state biopsy method is used to search out the molecular marker for identifying the benign and malignant pulmonary nodules, and the mass spectrometric detection and the subsequent bio-information analysis show that the molecular marker FGB provides a novel molecular target for the diagnosis of the pulmonary nodules and the observation of clinical effects.

Description

technical field [0001] The invention belongs to the technical field of cancer molecular diagnosis technology, and specifically relates to an exosome protein as a marker for differentiating benign and malignant pulmonary nodules and its application. Background technique [0002] Currently, the morbidity and mortality of lung cancer rank among the top among malignant tumors in the world. About 170,000 people die of lung cancer in the United States every year, accounting for 30% of the overall cancer deaths. With the rapid economic development and increasing air pollution, the incidence of lung cancer in my country is also showing a rising trend year by year. Low-dose CT is widely used in the screening of lung cancer. A large number of patients are diagnosed as pulmonary nodules by imaging every year, and most of them are confirmed to be benign pulmonary nodules. Therefore, it is necessary to correctly identify the discovered pulmonary nodules Its benign and malignant become ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6884
Inventor 匡牧宇孙艺华张辉标周子琅李多邓超
Owner HUADONG HOSPITAL
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