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RASL12 gene serving as diagnosis and treatment target of lung squamous cell carcinoma metastasis

A lung squamous cell carcinoma and genetic technology, applied in the biological field, can solve the problem of no obvious progress in the treatment of cancer patients

Active Publication Date: 2016-09-21
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there has been no significant progress in the treatment of cancer patients with extensive metastases or advanced metastases

Method used

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  • RASL12 gene serving as diagnosis and treatment target of lung squamous cell carcinoma metastasis
  • RASL12 gene serving as diagnosis and treatment target of lung squamous cell carcinoma metastasis
  • RASL12 gene serving as diagnosis and treatment target of lung squamous cell carcinoma metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Differential expression of embodiment 1 RASL12 gene

[0062] 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.

[0063] 2. Acquisition of RNA from lung squamous cell carcinoma metastasis tissue

[0064] 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 Nanodrop ND-1000 (A). The integrity of RNA was detected by electrophoresis on 1% formaldehyde denatured agarose gel and observed under ultravi...

Embodiment 2

[0073] Example 2 Differential expression of RASL12 protein

[0074] 1. Extract tissue total protein

[0075] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0076] 2. Western blot detection

[0077] The extracted protein was quantitatively subjected to SDS-PAGE electrophoresis, followed by membrane transfer, blocking, primary antibody incubation, secondary antibody incubation, and color development.

[0078] 3. Statistical processing

[0079]The gray value of the protein band was analyzed using Image J software, and the gray value of the target white 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 when P<0.05.

[0080] 4. Results

[0081] The result is as figure ...

Embodiment 3

[0082] Embodiment 3 RASL12 gene expression plasmid construction

[0083] 1. Construction of RASL12 gene expression vector

[0084] Amplification primers were designed according to the coding sequence of the RASL12 gene (as shown in SEQ ID NO.1). Amplify the coding sequence of the full-length RASL12 gene from the cDNA library of adult fetal brain (clontech company, article number: 638831), insert the above cDNA sequence into the eukaryotic cell expression vector pcDNA3.1, and connect the obtained recombinant vector pcDNA3.1 -RASL12 was used in subsequent experiments.

[0085] 2. Cell culture

[0086] The lung squamous cell line NCI-H520 was 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% fetal bovine serum at 37°C for 5 %CO 2 in the incubator.

[0087] Take NCI-H520 cells by 1×10 4 Inoculate / well into 24-well cell culture plates at 37°C, 5% CO 2 The cells were cultured in the incubato...

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Abstract

The invention discloses an RASL12 gene and an expression product thereof which can serve as molecular markers of diagnosis and treatment on lung squamous cell carcinoma metastasis. Whether a subject suffers from lung squamous cell carcinoma metastasis or not can be judged or whether the subject has the risk of suffering from lung squamous cell carcinoma or not can be diagnosed by detecting the content of the RASL12 gene and the expression product thereof in lung tissue of the subject. According to the RASL12 gene and the expression product thereof which can serve as the molecular markers of diagnosis and treatment on lung squamous cell carcinoma, it is found that RASL12 gene expression can inhibit adhesion, metastasis and invasion of lung squamous carcinoma cells by researching lung squamous carcinoma cells subjected to in vitro culture, and it is shown through research results that the RASL12 gene is a potential drug target for treating lung squamous cell carcinoma metastasis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of RASL12 gene in the diagnosis and treatment of lung squamous cell carcinoma metastasis. Background technique [0002] Tumor is currently one of the leading causes of death in the world, and more than 90% of cancer patients die from tumor metastasis. About 1 / 3 of breast cancer, colorectal cancer, cervical cancer, oropharyngeal cancer, and 1 / 4 of esophageal cancer, lung cancer, pancreatic cancer, bladder cancer, and gastric cancer patients have lymph node metastasis during surgery, which affects the patient's health. Treatment and Prognosis. With the aggravation of urban air pollution, lung cancer has become one of the most important lethal tumors in developed countries and cities. So far, there is still no significant progress in the treatment of cancer patients with extensive metastases or advanced metastases. Therefore, exploring the molecular mechanism of tumor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/00
Inventor 杨承刚边洋李海岩
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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