Spinal highly-metastatic human lung adenocarcinoma cell strain and construction method and application thereof

A technology of lung adenocarcinoma cell line and human lung adenocarcinoma, applied in the field of tumor biology, can solve the problem of lack of animal models of lung cancer spine metastasis, and achieve the effect of high spine metastasis rate

Active Publication Date: 2014-04-30
EAST CHINA NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main reason is the lack of reliable ani

Method used

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  • Spinal highly-metastatic human lung adenocarcinoma cell strain and construction method and application thereof
  • Spinal highly-metastatic human lung adenocarcinoma cell strain and construction method and application thereof
  • Spinal highly-metastatic human lung adenocarcinoma cell strain and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Construction of human lung adenocarcinoma cell line A549L6 with high metastases in the spine

[0060] Human lung adenocarcinoma cell line A549 cell line (ATCC No. CCL-185) cells were prepared according to 1.0×10 5 Each well was inoculated in a 24-well culture plate, and transfected with P6L4 plasmid (Promega Biotech Co.Ltd) the next day; the transfection steps and methods were carried out according to the instructions of Lipofectamin2000 reagent; after 24 hours of transfection, the medium was changed, and 600ug / ml G418 (purchased from German merck company) screened for three weeks to obtain positive cell clones; and the luciferase (Luciferase) fluorescence activity assay was performed on the obtained clones.

[0061] Human lung adenocarcinoma cell line A549 cell suspension 0.1ml (containing 1x10 5 cells) were injected into the left ventricle of nude mice, and the IVIS in vivo imaging system and X-ray imaging system were used to monitor the spinal metastasis ...

Embodiment 2

[0065] Example 2: Characteristic detection of human lung adenocarcinoma cell line A549L6 with high spinal metastases of the present invention

[0066] The cell line A549L6 of the present invention constructed in Example 1 is detected by routine series of experiments:

[0067] 1. After routine trypsinization and centrifugation of the cells, adjust the density to 1x10 6 / ml, 100ul / well was added to a black 96-well plate, and serially diluted, with 2 replicate wells for each gradient. Add 100ul of 300μg / ml luciferase substrate to each well, place it in an IVIS live imaging system (Caliper Life Sciences IVIS Lumina II, model: DS934N-BV-286) after 10 minutes, and the exposure time is 5 minutes, and the detection shows Both A549L0 and A549L6 stably expressed luciferase. Such as figure 1 As shown on the left, after multiple rounds of in vivo screening, when the number of cells is 1x10 6 , the fluorescence expression value of A549L6 was about 5.512 times that of A549L0; when the ...

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Abstract

The invention discloses spinal highly-metastatic human lung adenocarcinoma cell strain A549L6, the cell strain is preserved in China Center for Type Culture Collection (CCTCC) in April 25, 2013 with the accession number of CCTCC No:C201346. The invention also discloses a construction method and application of the spinal highly-metastatic human lung adenocarcinoma cell strain. The spinal highly-metastatic human lung adenocarcinoma cell strain is high in spinal metastatic rate, and can be widely applied to metastatic study models of lung adenocarcinoma, studies on metastatic mechanisms of the lung adenocarcinoma and screening and preparing of anti-metastasis drugs.

Description

technical field [0001] The invention relates to the technical field of tumor biology and the technical field of microbial animal cell lines, in particular to a human lung adenocarcinoma cell line with high metastases in the spine and its construction method and application. Background technique [0002] Cancer is currently the number one killer threatening human health. According to incomplete statistics from the World Health Organization, more than seven million patients die of cancer every year around the world. Lung cancer is one of the cancers with the highest morbidity and mortality in the world. About 170,000 people die from lung cancer in the United States every year, accounting for 30% of the total number of cancer deaths [1]. With the rapid economic growth in recent years, the incidence of lung cancer in my country has begun to double. About 400,000 people are diagnosed with lung cancer every year, and the incidence rate is as high as 61.4 / 100,000, making it the co...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/85A01K67/027C12Q1/02
Inventor 罗剑肖建如刘明耀蔡小攀
Owner EAST CHINA NORMAL UNIVERSITY
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