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Method for establishing inflammation-based pancreatic cancer animal model

A technique for establishing animal models and methods, applied in the field of establishing animal models of pancreatic cancer

Inactive Publication Date: 2015-11-11
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it has not been confirmed whether DBTC can induce pancreatitis in nude mice

Method used

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  • Method for establishing inflammation-based pancreatic cancer animal model
  • Method for establishing inflammation-based pancreatic cancer animal model

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Embodiment 1

[0087] A method for establishing an animal model of pancreatic cancer with an inflammatory basis, the steps are as follows:

[0088] The first step, the construction of pLVX-luciferase-IRES-ZsGreen1 plasmid:

[0089] 1) Prepare pLVX-IRES-ZsGreen1 digestion product and pGL3-Basic digestion product through the following reaction system and conditions,

[0090] reaction system:

[0091] DNA (pLVX-IRES-ZsGreen1, pGL3-Basic plasmids), 1 μg

[0092] 10×NEbuffer4 (+BSA), 3 μL

[0093] Endonuclease XhoI, 1 unit

[0094] Endonuclease XbaI, 1 unit

[0095] Add sterilized ultrapure water to a volume of 30 μL

[0096] Reaction conditions: 37°C water bath for 2h.

[0097] 2) Perform agarose gel electrophoresis on the digested product of pGL3-Basic, cut the gel and recover the gel block containing a 1700bp DNA fragment, and use the AxyPrepDNA Gel Recovery Kit to purify to obtain the luciferaseDNA fragment, which is gelled with AxyPrepDNA The specific steps for purification with the G...

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Abstract

The invention provides a method for establishing an inflammation-based pancreatic cancer animal model, which comprises the following steps: 1. constructing a pLVX-luciferase-IRES-ZsGreen1 plasmid; 2. establishing a slow-virus-mediated luciferase overexpression system; 3. establishing a pancreatic cancer cell line for expressing luciferase; 4. establishing a nude mouse pancreatic gland orthotopic transplantation tumor model; 5. establishing an inflammation model; and 6. establishing the inflammation-based pancreatic cancer animal model in the nude mouse body by continuing breeding the nude mouse for 30 days. The pancreatic cancer animal model established by the method can catalyze the substrate to emit light by using the luciferase, and the size of the tumor can be evaluated after the light is detected by the corresponding instrument and equipment.

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 pancreatic cancer with an inflammatory 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, significantly contribute to chronic tissue damage and subsequent neoplasia in these organs [1]. Chronic inflammation is closely related to the occurrence and development of pancreatic cancer [2]. The incidence of pancreatic cancer in the general population is approximately 0.007% [3], increasing to 4% in patients with chronic pancreatitis and as high as 40% in patients with hereditary pancreatitis [4]. The use of anti-inflammatory agents has been associated wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N5/10A01K67/027
Inventor 李伟吴梦瑶陶敏龚斐然
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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