Method for establishing colorectal cancer animal model with inflammation basis

A colorectal cancer, animal model technology, applied in the field of establishing colorectal cancer animal models

Inactive Publication Date: 2016-11-23
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Technical problem to be solved: Aiming at the shortcoming that there is no successful establishment of an animal model paradigm of inflammation on the growth and metastasis of colorectal cancer, the present invention provides a method for establishing an animal model of colorectal cancer with an inflammation basis

Method used

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  • Method for establishing colorectal cancer animal model with inflammation basis
  • Method for establishing colorectal cancer animal model with inflammation basis

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Embodiment 1

[0128] A method for establishing an animal model of colorectal cancer with an inflammatory basis, the steps are as follows:

[0129] The first step, the construction of pBabe-CMV-EGFP-puro plasmid:

[0130] 1) Prepare the pBabe-puro digestion product through the following reaction system and conditions, wherein the DNA is the pBabe-puro plasmid,

[0131] reaction system:

[0132] DNA, 1 μg

[0133] 10×NEbuffer3+BSA, 3 μL

[0134] Endonuclease BamHI, 1 unit

[0135] Endonuclease EcoRI, 1 unit

[0136] Add sterilized ultrapure water to volume 30 μL

[0137] Reaction conditions: 37°C water bath for 2h

[0138] 2) Perform agarose gel electrophoresis on the pBabe-puro digestion product, cut the gel and recover a DNA fragment containing 5100 bp in size, and use AxyPrep DNA Gel Extraction Kit for purification to obtain the purified pBabe-puro digestion product , wherein the specific steps for purification using the AxyPrep DNA Gel Recovery Kit are:

[0139] ① Cut off the agar...

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Abstract

The invention provides a method for establishing a colorectal cancer animal model with an inflammation basis. The method includes the following steps that firstly, pBabe-CMV-EGFP-puro plasmids are established; secondly, a retrovirus-mediated EGFP overexpression system is established; thirdly, a colorectal cancer cell line stably expressing EGFP is established; fourthly, an axillary colorectal cancer transplanted tumor model of group A nude mice is established; fifthly, an inflammation model of group B nude mice is established; sixthly, the colorectal cancer animal model with the inflammation basis is established, wherein an axillary tumor of the group A nude mice is excised and sheared into tumor blocks with the diameter of 2 mm or so through sterile eye scissors, one tumor block is taken and directly sewn to the wall of the colorectum of the group B nude mice, the fifth step is repeated, feeding is carried out for one week, the fifth step is repeated again, feeding continues for one week, and the cancer animal model is obtained. With the animal model established through the method, the existence of tumors can be verified, and the size of tumors can be estimated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for establishing an animal model of colorectal cancer with an inflammation basis. Background technique [0002] Epidemiological and clinical studies have confirmed that about 25% of adult tumors are caused by chronic inflammation. For example, chronic viral or bacterial infections of the stomach, liver, and cervix, as well as chronic exposure of the lungs to tobacco smoke or other environmental compounds, can significantly contribute to chronic tissue damage and subsequent tumorigenesis in these organs. The risk of colorectal cancer in chronic ulcerative colitis (Crohn's disease) is 4 to 20 times higher than that of normal people; the risk of malignant transformation in hemorrhagic ulcerative colitis is greater, and 50% of those with a course of more than 10 years will develop into cancer; The use of inflammatory agents was associated with a reduction in the inc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N15/65C12N5/10A01K67/027
CPCA01K67/027A01K2207/12A01K2207/20A01K2227/105A01K2267/03A01K2267/0331C07K14/43595C12N15/65C12N15/86C12N2740/10043
Inventor 李伟周进吴梦瑶支巧明龚斐然
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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