Mouse esophageal cancer model and construction method thereof

A method for establishing esophageal cancer technology, which is applied in the field of biomedicine, can solve the problems of complex cell components, esophageal cancer cell lines are rarely used, and the applicability of rat tumor cell lines is not good enough to achieve good proliferation effect

Active Publication Date: 2019-11-08
SHENZHEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the establishment system of this cell line, metastatic tumor tissue was used instead of esophageal tissue in situ, and there were no clear indicators in the later stage to determine the origin of the established cell line from the esophagus; therefore, it was difficult to identify the established cell line as esophageal cancer cell line
At the same time, the cell lineage method used in this model only retains the largest clones, which may not be able to show the heterogeneity of tumor cells to the greatest extent
In addition, in the largest clone obtained by tissue culture, the cell components may be relatively complex, and it will take a long time to culture in the later stage to achieve the purpose of purifying and enriching tumor epithelial cells
Although this model established rat esophageal cancer cell lines in 2007, until now, these rat esophageal cancer cell lines have been rarely used in research
In addition to some of the above shortcomings, the applicability of rat tumor cell lines is not as good as that of mouse tumor cell lines

Method used

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  • Mouse esophageal cancer model and construction method thereof
  • Mouse esophageal cancer model and construction method thereof
  • Mouse esophageal cancer model and construction method thereof

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Experimental program
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Effect test

Embodiment 1

[0045] The establishment method of mouse esophageal cancer model of the present invention is as follows:

[0046] (1) Construction of primary mouse esophageal cancer model

[0047] 4-Nitroquinoline-1-oxide was formulated into a 5mg / ml stock solution using 1,2-propanediol, and then formulated into a 100ug / ml 4NQO aqueous solution using sterile water. The 4NQO aqueous solution was divided into drinking bottles for mice, and the water bottles were wrapped with tinfoil and placed in mouse cages for mice to drink. Regularly pay attention to the remaining amount of solution in the water bottle, and add 100ug / ml 4NQO aqueous solution at any time. After the mice continued to drink sterile water containing 100ug / ml 4NQO for 16 weeks, they switched to drinking pure sterile water; after drinking pure sterile water for 12 weeks, the mice were sacrificed according to animal ethics requirements, and the esophagus was cut off in a safety cabinet , observe the formation of carcinoma in situ...

Embodiment 2

[0054] 1. Identification of molecular biological characteristics of mouse esophageal cancer cell line mEC25

[0055] The epithelial and tumorigenic properties of the cell line mEC25 cultured in 10% FBS-DMEM for more than 10 passages were identified. mEC25 cells were grown adherently on polylysine-treated coverslips, then fixed with 4% neutral formaldehyde for 10 minutes and permeabilized with 0.01% triton X-100 for 10 minutes at room temperature, and incubated with 10 After blocking with % sheep serum for 30 minutes, use pan-cytokeratin, E-cadherin, β-catenin, SOX2 and P63 antibodies respectively, each antibody concentration is 10ug / ml, incubate at room temperature for 2 hours, after washing, use Alexa Fluor 594-conjugated Goat anti-Rabbit (β-catenin, SOX2 and P63) or Alexa Fluor 594-conjugated Goat anti-mouse (pan-cytokeratin and E-cadherin) were incubated at room temperature for 1 hour. The nuclei were stained with Hoechst33342 (2ug / ml, 10min), and the immunofluorescence re...

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Abstract

The invention relates to the technical field of biomedicine, in particular to a mouse esophageal cancer model and a construction method thereof. The construction method includes: inducing mice by a chemical carcinogenic agent to construct a primary mouse esophageal cancer model to obtain an esophageal cancer in-situ tumor tissue; transplanting the esophageal cancer in-situ tumor tissue to naked mice subcutaneously, and performing in-vivo culture to obtain a transplanted tumor tissue; sequentially subjecting the transplanted tumor tissue to removal of part of fiber cells according to a rapid adherence method, enriching of epithelial tumor cells according to an epithelial culture method and removal of residual fiber cells according to a rapid digestion method, and separating to obtain the mouse esophageal cancer model. The mouse esophageal cancer model used for rapid construction of perfectly-immune mouse esophageal cancer model is constructed, and the stable and reliable mouse esophageal cancer model construction method is formed, so that a gap of the mouse esophageal cancer model is filled, current related problems caused by lack of the mouse esophageal cancer model is solved, anda suitable research tool is provided for tumor immunology research of esophageal cancers.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a mouse esophagus cancer model and its establishment method. Background technique [0002] my country is a country with high morbidity and mortality of esophageal cancer in the world, accounting for more than 50% of the world's total. There are about 250,000 newly diagnosed cases of esophageal cancer every year. Esophageal cancer seriously endangers the life and health of Chinese people; Cystoid cell carcinoma is the most common type of esophageal cancer. The treatment of esophageal squamous cell carcinoma is still dominated by traditional chemotherapy, radiotherapy and surgery, and the cure rate of advanced patients is low. Immunotherapy has demonstrated its great potential in melanoma, lung cancer and other cancers; however, in the field of esophageal cancer research, there are few related studies on immunotherapy. One of the most important reasons is that the current rese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C12N5/09
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0331C12N5/0693
Inventor 付利黄土雄陈燕雯
Owner SHENZHEN UNIV
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