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Method for inducing vicious transformation of human hepatocytes by trichloroethylene and application of method

A trichloroethylene and hepatocyte technology, applied in the field of cell biology, can solve the problem of few different cell lines, eliminate species differences, facilitate signal pathways and early molecular effects, and be suitable for large-scale culture.

Pending Publication Date: 2022-08-02
安徽理工大学环境友好材料与职业健康研究院(芜湖) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] According to the description of the patent application number CN201410842795.0, the use of benzo(a)pyrene can induce malignant transformation of cells, but currently there are few different cell lines that trichlorethylene can use, and it is urgent to supplement

Method used

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  • Method for inducing vicious transformation of human hepatocytes by trichloroethylene and application of method
  • Method for inducing vicious transformation of human hepatocytes by trichloroethylene and application of method
  • Method for inducing vicious transformation of human hepatocytes by trichloroethylene and application of method

Examples

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Effect test

Embodiment 1

[0093] Example 1. Screening of cells of the present invention

[0094] Take the normal human liver cell line THLE-2 (ATCC), add 2mM trichloroethylene to the medium for long-term culture, observe the cell growth status under the microscope every 2-3 days, including cell morphology, survival rate, growth rate Wait. The study found that the continuous stimulation of 2mM trichloroethylene induced significant changes in cell morphology, but the cell viability did not decrease, and it also showed stronger growth ability. This phenotypic change persisted after the removal of trichloroethylene, indicating that the The cells have been irreversibly changed and transformed into a new cell type with stable properties, which is the cell line obtained by the screening of the present invention, which is named "human normal hepatocyte trichloroethylene cell line THLE-2-TCE".

[0095]The cells obtained by this screening were taken for cryopreservation, and were recovered, cultured and cryopre...

Embodiment 2

[0096] Example 2. Morphological changes of cells obtained by screening

[0097] Comparing the cell line obtained by the aforementioned screening with the normal liver cells not treated with trichloroethylene, it is found that the morphology of the cell line of the present invention has changed significantly. like figure 1 As shown, when THLE-2 was transformed for 30 generations, the cell morphology changed significantly, and the cells were pleomorphic under the microscope, the cells grew in no direction, and the arrangement was disordered. Agglomerate growth, disorderly arrangement, and many cells are burr-like around. After 30 passages of cells covered the bottom of the petri dish, there was overlapping growth, the cell body became larger, the cell outline was gradually enhanced, and multinucleated giant cells were seen. This morphological change indicates that the cell has exhibited cellular characteristics different from the source cell and is caused by continuous stimula...

Embodiment 3

[0099] Example 3. Growth of cells obtained by screening

[0100] The cell lines obtained by the aforementioned screening were compared with normal hepatic cells not treated with trichloroethylene. The result is as figure 2 , it was found that the growth rate of the cell line of the present invention was significantly increased, indicating that compared with the source cell, the cell line obtained by screening had an abnormally fast growth ability, indicating that the continuous stimulation of trichloroethylene changed the cell division and proliferation ability.

[0101] The above results show that the cell line obtained by the screening of the present invention can be used as a new cell model for studying how trichloroethylene affects cell division and proliferation.

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Abstract

The invention discloses a technology for establishing a human normal hepatocyte in-vitro trichloroethylene induced malignant transformation model. The technology comprises the following steps: firstly, inducing human normal hepatocyte in-vitro malignant transformation in vitro; and then identifying the in-vitro malignant transformation of the human normal hepatocytes. According to the technical scheme, a new experimental model is provided for better knowing the effect and biological characteristics of the trichloroethylene for inducing the malignant transformation of human normal hepatocytes and deeply researching the pathogenic mechanism of the trichloroethylene or screening medicines for intervening the malignant transformation of the hepatocytes.

Description

technical field [0001] The present invention relates to the field of cell biology, and more particularly, the present invention relates to a technical method for inducing malignant transformation of hepatocytes by industrial chemicals. Background technique [0002] With the development of medical and toxicology research technology, the traditional toxicology evaluation mode that mainly relies on animal in vivo experiments has changed, and in vitro model research has become one of the important directions of toxicology development. Traditional toxicology research methods are mainly based on animal in vivo experiments, with high cost and long experimental period, which are not conducive to high-throughput screening. Uncertainties related to humans lead to large deviations, and the observation end point is clinical characteristics or pathological changes, which cannot be detected. Early changes in vivo make it difficult to explain toxicity mechanisms from the perspective of sig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071C12Q1/02
CPCC12N5/0693C12N5/067G01N33/5014C12N2501/999
Inventor 唐小龙曹伟娅周淑萍张殷慈李阿敏马咏芳徐如月
Owner 安徽理工大学环境友好材料与职业健康研究院(芜湖)
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