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Model cell strain for malignant transformation of lung cell induced by nano-silica dioxide and application thereof

A technology of model cells and cell lines, applied in the field of cell engineering, can solve problems such as undefined occupational exposure limits and insufficient carcinogenicity

Inactive Publication Date: 2019-01-29
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, currently the nano-SiO 2 The research on the harmful biological effects of chlorophyll is still in its infancy, and the research on its carcinogenicity is not deep enough, and there is no definition of the occupational exposure limit

Method used

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  • Model cell strain for malignant transformation of lung cell induced by nano-silica dioxide and application thereof
  • Model cell strain for malignant transformation of lung cell induced by nano-silica dioxide and application thereof
  • Model cell strain for malignant transformation of lung cell induced by nano-silica dioxide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 The process of model establishment

[0024] 1. Cell selection (Shanghai) human lung bronchial epithelial cells Beas-2B cells from ATCC cell bank;

[0025] 2. After the cells are purchased, STR typing and identification will be carried out by Readwei Gene Company, and the identification results are completely consistent with the Beas-2B of the ATCC cell bank;

[0026] 3. Dosage selection

[0027] 1) Beas-2B cells were cultured, and the culture conditions were fetal bovine serum from Sciencell, DMEM high-glucose medium from hyclone, 5% CO 2 ,37 degree;

[0028] 2) The cells were cultured and passaged for 3 times, and then inoculated in a 96-well plate with 10,000 cells per well. 24h after inoculation, nano-SiO 2 poison;

[0029] 3) Poisoning process: firstly discard the cell culture medium, wash the cells three times with PBS buffer, and then carry out the poisoning. Seven dose groups are set, 0 μg / ml, 2.5 μg / ml, 5 μg / ml, 10 μg / ml, respectively. ml, 25μg / ml...

Embodiment 2

[0039] Example 2 Tumor-bearing experiment in nude mice

[0040] 1) Beas-P40 cells and Beas-2B SiO 2 P40 cells were cultured with fetal bovine serum from Sciencell, DMEM high-glucose medium from hyclone, 5% CO2, 37°C;

[0041] 2) Beas-P40 cells and Beas-2B SiO 2 P40 cells were cultured and passaged, and the cells were grown to log phase;

[0042] 3) Beas-P40 cells and Beas-2B SiO 2 P40 cells were collected, centrifuged at 1200 rpm for 3 min, resuspended in culture medium, counted, and adjusted to a cell concentration of 5 × 10 7 Cells / ml, 200 μl of cells were inoculated into the armpits of nude mice (purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.);

[0043] 4) Observing regularly after inoculation, measuring the size of the tumor of the nude mice, and finding that the tumor gradually increased, 24 days after the inoculation, the nude mice were killed by dislocation, the size and weight of the tumor were measured, and pathological detection was c...

Embodiment 3

[0045] Example 3 Gene chip detection

[0046] 1) Beas-P40 cells and Beas-2B SiO 2 P40 cells were cultured with fetal bovine serum from Sciencell, DMEM high-glucose medium from hyclone, 5% CO 2 ,37 degree;

[0047] 2) Beas-P40 cells and Beas-2B SiO 2 P40 cells were cultured and passaged. After 3 passages, the cells were collected after the cells had grown to the logarithmic phase, and 3 dishes were collected in each group and frozen at -80 degrees;

[0048] 3) It is stored and transported on dry ice, and sent to Shanghai Qiming Biotechnology Co., Ltd., and the Affymetrix ZebrafishGene 1.0ST chip is used for cell gene expression profile detection and bioinformatics analysis;

[0049] 4) The analysis of the global signal transduction network showed that the tumor suppressor gene p53 was located at the core of the global signal transduction network, indicating that the p53 gene played a key role in the malignant transformation of cells induced by nano-silica.

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Abstract

The invention provides a model cell strain for malignant transformation of a lung cell induced by nano-silica dioxide and an application thereof. The accession number of the model cell strain is CGMCCNO. 12676. The model cell strain and a daughter cell strain thereof can provide a basis for carcinogenicity research by utilizing in vitro experiments, can be used to study a mechanism of revealing carcinogenicity of the nano-silica dioxide, and can also be used for screening early biomarkers of carcinogenesis of the nano-silica dioxide and developing corresponding detection kits and drugs related to lung cancer treatment.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and in particular relates to nano-SiO 2 Model cell line for malignant transformation of lung cells and its application. Background technique [0002] Nano silica (silica dioxide, SiO 2 ) is considered to be one of the main components of the inert core of atmospheric particulate matter. Recent studies have shown that both environmental nanoparticles and engineered nanomaterials inevitably enter the ecosystem and atmospheric environment, and nano-SiO 2 It is one of the most important ingredients. SiO 2 As a crustal component, the average content in the soil is 57.8% to 64.8%, so nano-SiO 2 As a natural nanomaterial, it can enter the atmosphere through soil dust, coal soot, dust, etc. Meanwhile, nano-SiO 2 It is also one of the most commonly used engineered nanomaterials in the world, and is widely used in the fields of materials, cosmetics and biomedicine. Among the nanomaterial co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12Q1/02A61P35/00
CPCC12N5/0688A61K35/42C12N2503/00G01N33/5044
Inventor 李艳博孙志伟郭彩霞周显青
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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