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Construction method of in-situ liver cancer model of SD rats (Sprague-Dawley) having high liver hardness background, and application thereof

A construction method and technology of rat liver cancer cells, applied in the field of construction of SD rat orthotopic liver cancer model with high liver stiffness background, can solve the problems of high cost, long modeling period, and difficulty in obtaining Buffalo rats, and achieve good economical results. Benefit and application value, low cost, and short tumor formation time

Active Publication Date: 2022-02-11
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0014] The first purpose of the present invention is to provide a method for constructing an orthotopic liver cancer model in SD rats with a high liver stiffness background, which can better simulate and reproduce the clinical pathological and biochemical characteristics of patients with liver cirrhosis and liver cancer, and solve the problem of high cost, difficulty in obtaining Buffalo rats and modeling Long period, relatively complex characteristics and other issues, to form a more practical and efficient new alternative animal model

Method used

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  • Construction method of in-situ liver cancer model of SD rats (Sprague-Dawley) having high liver hardness background, and application thereof
  • Construction method of in-situ liver cancer model of SD rats (Sprague-Dawley) having high liver hardness background, and application thereof
  • Construction method of in-situ liver cancer model of SD rats (Sprague-Dawley) having high liver hardness background, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] as per figure 1 The method shown is to construct an orthotopic liver cancer model in SD rats with high liver stiffness background, and the steps are as follows:

[0043] The first step, inducing SD rats with high liver stiffness:

[0044] Male SD rats aged 4 to 6 weeks were selected, and SD rats were injected with carbon tetrachloride subcutaneously in the abdomen. 1), 2ml / kg, 2 times a week, 12 weeks in total, the condition of rat liver after induction is as follows figure 2 As shown in A.

[0045] The second step is to establish an orthotopic liver cancer model in SD rats with high liver stiffness background:

[0046] Cells: expansion of rat liver cancer cells McA-RH7777;

[0047] Glucocorticoid pretreatment: Intramuscular injection of dexamethasone at a dose of 5 mg / kg for 5 days, starting from 2 days before orthotopic liver cancer implantation in rats until the 2nd day after operation (d-2~d2);

[0048] Preoperative preparation: anesthetized with 2% sodium pen...

Embodiment 2

[0051] To verify the orthotopic liver cancer model of SD rats with high liver stiffness background in Example 1, the steps are as follows: evaluate the liver stiffness of the previously formed SD rats with high liver stiffness, and randomly select 6 SD rats with high liver stiffness to perform liver elasticity ultrasound detection. The results suggest that the average liver hardness value of SD rats with high liver hardness is 17.03KPa (Table 1), which can better simulate the liver hardness value of human liver cirrhosis.

[0052] Table 1: Liver elastic ultrasound hardness value of SD rat orthotopic liver cancer model with high liver stiffness background

[0053] case 1 2 3 4 5 6 mean Stiffness value(KPa) 15.45 19.68 15.02 17.67 17.85 16.54 17.03

[0054] The serum liver function indexes of the orthotopic liver cancer model of SD rats with high liver stiffness were detected, and it was found that the liver enzymes (ALT, AST) of SD rats in the hi...

Embodiment 3

[0059] In this example, the orthotopic liver cancer model of SD rats with high liver stiffness background was used to conduct relevant in vivo and in vitro research experiments on the influence of the stromal microenvironment on liver cancer. The steps are as follows:

[0060] Using the above method to construct rat liver cirrhosis model, prepare liver cirrhosis rats and divide them into four groups: ①cirrhosis rats, WT; ②cirrhosis rats, NC; ③liver cirrhosis rats, shITGB1; ④liver cirrhosis rats, shPiezo1.

[0061] Rat liver cancer cell McA-RH777 was cultured in a culture flask, and the cells were collected in the logarithmic proliferation phase, washed with PBS, counted on a counting plate, and made into 1.6×10 6 Rat liver cancer cell / 100ul cell suspension, add 100ul Marigel to mix by pipetting to avoid air bubbles.

[0062] According to the above method, the rats in each group were implanted with cells under the liver capsule, and the rats were sacrificed 2 weeks later, and ...

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Abstract

The invention discloses a construction method of an in-situ liver cancer model of SD rats (Sprague-Dawley) having a high liver hardness background, and an application thereof. The construction method comprises the following steps: performing hypodermic injection of carbon tetrachloride on the abdomen of SD rats to induce the formation of high liver hardness on the SD rats, injecting Buffalo rat liver cancer cells mixed with low-growth factor Matrigel under a liver envelope of the SD rats, performing short-term intramuscular injection in combination with a small dose of glucocorticoid , and forming an in-situ liver cancer model of the SD rats having a high liver hardness background after 12-14 days. SD rats and rat liver cancer cells adopted by the method are easy to obtain and low in cost, and have the advantages of high tumor formation rate, controllable tumor number and position, short tumor formation time, high metastasis and the like, so that the problems of technical difficulty and modeling cost of the establishment of a hardness background liver cancer animal model in the prior art are effectively solved, malignant pathological characteristics of liver cancer patients having a high liver hardness background are well simulated, and a practical and efficient novel liver cancer animal model is provided for analysis of mechanisms such as gene functions, radiotherapy resistance, and immunosuppression in a hardness mechanical micro-environment.

Description

technical field [0001] The invention relates to the technical field of animal models, in particular to a method for constructing an orthotopic liver cancer model in SD rats with high liver stiffness and its application. Background technique [0002] Matrix hardening is mainly caused by excessive deposition and cross-linking of extracellular matrix proteins. This change in mechanical hardness often destroys the balance of cell surface forces, promotes the aggregation of integrins and the formation of focal adhesions, and transforms the mechanical signal of exogenous hardness into the cell. Biochemical signals, thereby affecting and changing cell biological phenotypes and characteristics. Matrix sclerosis is a typical biomechanical phenotype of solid tumors, often accompanied by tumor occurrence and progression. Currently, matrix stiffness assessment (elastic ultrasound imaging) has been used as a method for predicting the development and prognosis of solid tumors such as brea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02A61D1/00A61D7/00
CPCA01K67/02A61D1/00A61D7/00A01K2267/03A01K2227/105
Inventor 李苗王咪咪张希崔杰峰陈荣新任正刚
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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