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Molecular marker differentiating metastatic squamous cell lung carcinoma from non-metastatic squamous cell lung carcinoma

A technology of lung squamous cell carcinoma and product, applied in the field of diagnosis and treatment of lung squamous cell carcinoma metastasis by TEX13B gene, which can solve the problem of unclear metastasis mechanism and so on.

Inactive Publication Date: 2016-07-13
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite many studies on lung cancer metastasis, the mechanism of metastasis remains unclear

Method used

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  • Molecular marker differentiating metastatic squamous cell lung carcinoma from non-metastatic squamous cell lung carcinoma
  • Molecular marker differentiating metastatic squamous cell lung carcinoma from non-metastatic squamous cell lung carcinoma
  • Molecular marker differentiating metastatic squamous cell lung carcinoma from non-metastatic squamous cell lung carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Differential expression of embodiment 1 TEX13B gene

[0063] 1. Sample acquisition: 40 cases of lung squamous cell carcinoma tissues (including 20 samples with metastasis and 20 samples without metastasis). The above samples are surgical resection specimens of patients with lung squamous cell carcinoma. All the above specimens were obtained with the consent of the organizational ethics committee. The clinical data of the tissue samples include: gender, age, tumor size, pathological grade (Edmonson), metastasis or recurrence, etc.

[0064] 2. Acquisition of RNA from lung squamous cell carcinoma metastasis tissue

[0065] The total RNA of metastatic lung squamous cell carcinoma tissue was extracted by Trizol one-step method, and the purity of the RNA solution was determined by reading the absorbance values ​​at 260nm and 280nm by NanodropND-1000 (A). The integrity of RNA was detected by electrophoresis on 1% formaldehyde denatured agarose gel and observed under ultravio...

Embodiment 2

[0074] Differential expression of embodiment 2 TEX13B protein

[0075] 1. The research object is the same as in Example 1.

[0076] 2. Extract tissue total protein

[0077] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0078] 3. Western blot detection

[0079] The extracted protein was quantitatively subjected to SDS-PAGE electrophoresis, followed by membrane transfer, blocking, primary antibody incubation, secondary antibody incubation, and color development.

[0080] 4. Statistical processing

[0081] The gray value of the protein band was analyzed using ImageJ software, and the gray value of the TEX13B protein band was normalized with β-actin as an internal reference. The results and data are expressed in the form of mean ± standard deviation, and SPSS13.0 statistical software is used for statistical analysis. The difference between the two is tested by t test, and it is considered statistically significant whe...

Embodiment 3

[0084] Example 3 Expression of TEX13B gene in lung squamous cell carcinoma cell lines

[0085] 1. Cell culture

[0086] The lung squamous cell line NCI-H520, NCI-H596, NCI-H2170, NCI-H226 were cultured in DMEM medium and 10% fetal bovine serum, and the human lung epithelial cell line BEAS-2B was cultured in BEGM medium and 10% Cultured in fetal bovine serum at 37°C, 5% CO 2 in the incubator.

[0087] 2. QPCR

[0088] 2.1 Extraction of total cellular RNA: Extraction of total cellular RNA was performed using the RNA extraction kit from QINGEN Company, and performed according to the instruction manual.

[0089] 2.2 Reverse transcription

[0090] The reverse transcription of RNA was carried out using the reverse transcription kit of TAKARA company.

[0091] 2.3 QPCR

[0092] (1) Primer design

[0093] QPCR amplification primers were designed according to the coding sequences of TEX13B gene and GAPDH gene in Genbank, and synthesized by Shanghai Sangon Bioengineering Technolo...

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Abstract

The invention discloses application of TEX13B gene and an expression product thereof to preparation of diagnostic tools for squamous cell lung carcinoma metastasis. According to the invention, a gene chip and a QPCR experiment prove that the TEX13B gene has difference in the expression of squamous cell lung carcinoma non-metastatic samples and squamous cell lung carcinoma metastatic samples, so that the TEX13B gene is believed to be a molecular marker for diagnosing squamous cell lung carcinoma metastasis. A culture in vitro cell experiment proves that restraining the expression of the TEX13B gene can restrain the adhesion, migration and invasion of lung squamous carcinoma, so that the TEX13B gene is believed to be a drug target for treating squamous cell lung carcinoma metastasis. As a new molecular marker, the TEX13B gene and the expression product thereof have a wide clinical application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of TEX13B gene in the diagnosis and treatment of lung squamous cell carcinoma metastasis. Background technique [0002] According to histological classification, lung cancer is generally divided into four types: squamous cell carcinoma, adenocarcinoma, large cell carcinoma, and small cell carcinoma. Among them, squamous cell carcinoma is the most common, accounting for about 40%-70% of all lung cancers. Lung cancer is one of the most common malignant tumors, and its mortality rate ranks first among human malignant tumors. In most countries of the world, lung cancer ranks first among common malignant tumors in men, accounting for 38.03% of male malignant tumor deaths; in women, its mortality rate has surpassed breast cancer to become the primary cause of tumor death. In recent years, due to the adoption of multidisciplinary comprehensive treatment based on surgery, th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00C12Q2600/158G01N33/57423G01N33/57484G01N33/68G01N2333/47
Inventor 杨承刚李曙光肖武
Owner BEIJING MEDINTELL BIOMED CO LTD
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