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Application of genes VSIG10L as lung squamous carcinoma metastasis diagnosis and therapy marker

A lung squamous cell carcinoma and genetic technology, which is applied in the determination/testing of microorganisms, medical preparations containing active ingredients, biological testing, etc., can solve the problems of time-consuming, increased patient suffering, inability to detect subclinical lesions and recessive lesions. Micrometastases and other issues

Active Publication Date: 2016-06-08
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these imaging data play an important role in diagnosis, there are also many limitations, such as a high false positive rate, and the inability to detect occult lesions, subclinical lesions, and micrometastases
In addition, the diagnostic methods for lung cancer also include invasive operations such as bronchoscopy and puncture biopsy. Such methods are time-consuming and increase the suffering of patients.

Method used

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  • Application of genes VSIG10L as lung squamous carcinoma metastasis diagnosis and therapy marker
  • Application of genes VSIG10L as lung squamous carcinoma metastasis diagnosis and therapy marker
  • Application of genes VSIG10L as lung squamous carcinoma metastasis diagnosis and therapy marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Differential expression of embodiment 1 VSIG10L 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 ultravi...

Embodiment 2

[0074] Differential expression of embodiment 2VSIG10L 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 β-actin was used as an internal reference to normalize the gray value of the VSIG10L protein band. 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 ...

Embodiment 3

[0084] Example 3 Expression of VSIG10L 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 VSIG10L gene and GAPDH gene in Genbank, and synthesized by Shanghai Sangon Bioengineering Techno...

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Abstract

The invention discloses a gene marker. The gene marker VSIG10L can be used for judging whether metastasis of lung squamous carcinoma occurs or not or pre-judging lung squamous carcinoma metastasis risks. The gene marker VSIG10L further can be used for preparing medicines for inhibiting or preventing lung squamous carcinoma metastasis. The gene marker has the advantages that novel diagnosis methods can be provided for clinically diagnosing lung squamous carcinoma metastasis at molecular level, and novel medicine targets can be provided for gene therapy on lung squamous carcinoma metastasis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of VSIG10L gene in the diagnosis and treatment of lung squamous cell carcinoma metastasis. Background technique [0002] Lung cancer is a major disease that seriously threatens human life and health. In 2011, the World Health Organization (WHO) International Cancer Research Agency IARC released the results of 2008 global statistical data, showing that lung cancer is the malignant tumor with the highest incidence and mortality. Every year, there are 1.608 million new cases and 1.377 million deaths worldwide, accounting for 12.7% and 18.2% of all malignant tumors. According to statistics released by the American Cancer Society, it is estimated that in 2012, the number of new lung cancer cases in the United States will reach 226,000, and the number of deaths due to lung cancer will reach 160,000. In my country, the results of the third national retrospective sampling survey on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/02
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57423G01N33/57484G01N33/68G01N2333/47
Inventor 杨承刚孙耀兰林慧芳
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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