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Method for constructing oral squamous cell carcinoma animal model

An animal model, oral mucosal technology, applied in biochemical equipment and methods, teaching models, animal cells, etc., can solve problems such as poor repeatability, systematic research on biological characteristics, and inability to simulate the natural process of tumorigenesis. The effect of less differences in biological characteristics and stable biological characteristics

Inactive Publication Date: 2009-07-01
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The existing technology has the following disadvantages: the incidence of spontaneous animal tumor models is extremely low, and the reproducibility is also poor
The disadvantages of xenogeneic animal tumor models established by human tumor cell lines are that they cannot well simulate the natural occurrence of tumors, and the biological characteristics of tumors change greatly after xenogeneic inoculation
Animal oral tumors and cell lines induced by chemical carcinogens can simulate the natural process of tumorigenesis, but there are only a few cell lines abroad, and no systematic research has been done on their biological characteristics. The biological characteristics of the cell lines are unstable. Difficult to use routinely as a cell line model

Method used

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  • Method for constructing oral squamous cell carcinoma animal model
  • Method for constructing oral squamous cell carcinoma animal model
  • Method for constructing oral squamous cell carcinoma animal model

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0027] Experimental Example 1 Tumor tissue and cell culture

[0028] 1. Materials: cytokeratin (cytokeratin, CK) monoclonal antibody (clone AE1 / AE3) and vimentin (vimentin, Vim) monoclonal antibody were purchased from DAKO Company, and the working concentration was 1:200 dilution. 4-nitroquinoline-1-oxide (4-nitroquinoline-1-oxide, 4NQO) was purchased from Sigma Company; Cycle TEST TM PLUS DNA kit was purchased from BD Company, USA. Female SD rats and Nu / nu nude mice were purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences.

[0029] 2. Method

[0030] 1) Add 4-nitroquinoline-1-oxide (4-nitroquinoline-1-oxide, 4NQO) to the drinking water to make the final concentration 0.02 g / L; feed 80 SD rats. After observation and feeding for 36 weeks, a total of 61 survived and had tumor formation. Oral mucosal epithelial tumor tissues were excised under aseptic conditions, and those confirmed as malignant oral mucosal tumors by quick frozen section were cult...

experiment example 2

[0032] Experimental example 2 Identification results of biological characteristics of cells grown in vitro

[0033] 1. Materials and main instruments: Cytokeratin (cytokeratin, CK) monoclonal antibody (clone AE1 / AE3) and vimentin (vimentin, Vim) monoclonal antibody were purchased from DAKO Company, and the working concentration was 1:200 dilution. Biological inverted microscope (TE300, Japan NICKON company); transmission electron microscope (transmission electron microscope, TEM, CM-120 type transmission electron microscope, Netherlands PHILIP company); scanning electron microscope (scanning electron microscope, SEM; QUANTA-200 environmental scanning electron microscope, Netherlands PHILIP company) company). Flow cytometer (FACSCalibur, American BD Company).

[0034] 2. Method

[0035] 1) Use phase contrast and differential interference inverted biological microscope to observe the growth status and shape of cells: Rca-T cells are mostly polygonal or spindle-shaped epithelio...

experiment example 3

[0048] Experimental Example 3 Subcutaneous Inoculation of Nude Mice Tumor Formation and Experimental Lung Metastasis Results

[0049] 1) Subcutaneous inoculation tumor formation experiments showed that: Rca-T and Rca-B cell lines were inoculated subcutaneously in nude mice with 3×10 6 The cells were sacrificed after 20 days, and tumors were formed in 16 subcutaneous injection points of each cell line, and the tumor formation rate was 100%. Observation of subcutaneous tumor formation tissue by pathological section of all tumor tissues showed a typical structure of squamous cell carcinoma, large cell atypia, multinucleated tumor giant cells and pathological mitosis were common, and cancer nests could be seen in the subcutaneous tumor formation tissue, which can be seen to keratinized beads (see Figure 17 ).

[0050] 2) The results of experimental lung metastasis experiment: Rca-T and Rca-B cells were inoculated into the tail vein of nude mice at 3×10 6 21 days after inoculat...

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Abstract

The invention relates to a method for constructing animal models, in particular to a method for constructing oral mucosa carcinoma animal models. The method adopts the technical proposal that the method for constructing the oral mucosa carcinoma animal models comprises the following steps: firstly, culturing a tumor tissue and tumor cells; and identifying biological characteristics of in vitro growth of the cells. The method for constructing the oral mucosa carcinoma animal models has the advantages that: firstly, Rca-B cell lines and Rca-T cell lines constructed by the method all come from single cloned tumor cells induced by SD pure mouse chemical carcinogens, and have the same genetic background and stable biological characteristics; and secondly, the characteristics of the cell lines such as hypodermal tumor formation in a nude mouse and high experimental liver transfer rate provide two practical animal models for study on prevention and treatment of oral mucosa epicytoma.

Description

technical field [0001] The invention relates to a method for constructing an animal model, in particular to a method for constructing an animal model of oral mucosal cancer. Background technique [0002] Oral mucosal epithelial squamous cell carcinoma is the most common type of malignant tumor in the oral and maxillofacial region. The 5-year survival rate of patients after comprehensive treatment based on surgery hovers around 50%. Establishing a cell model with stable biological properties and the ability to form transplanted tumors in vivo to simulate the occurrence and biological characteristics of human oral squamous cell carcinoma is a necessary condition for preclinical research on the basis, diagnosis and treatment of oral squamous cell carcinoma. At present, there are mainly four methods for establishing oral tumor cell lines and their tumor-bearing animal models at home and abroad: 1. Spontaneous animal tumor models and cell lines, which occur naturally in experime...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06G09B23/36C12N5/09
Inventor 陈万涛邱蔚六蒋灿华周晓健冯元勇张萍万澎波叶冬霞徐骎刘京苏严明张燕
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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