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Method of establishing animal model of HER2 positive tumor by using normal mouse

An animal model and mouse technology, applied in the biological field, can solve the problems of animal death, high cost, cumbersome operation, etc., and achieve the effects of wide application prospects, saving test costs, and overcoming limitations

Pending Publication Date: 2017-09-08
LINK HEALTH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Several existing animal models have their limitations, and it is difficult to truly evaluate the efficacy of tumor vaccines: ①The traditional immunodeficiency animals (nude mice or SCID mice) used to evaluate the efficacy of antitumor drugs, due to the deficiency of the immune system , can not be used in the evaluation of the efficacy of tumor vaccines, because tumor vaccines achieve anti-tumor effects by activating the immune system of the body; Immunosuppressive, human tumor cells can be inoculated, but immunosuppressed mice are not suitable for evaluating the efficacy of tumor vaccines; ③ immune reconstruction mice can rebuild the immune system in immunodeficient animals, but the operation is cumbersome and the stability is not good. And animals are prone to die due to immune rejection; ④ Transgenic mice are currently the most suitable animal models for evaluating the efficacy of tumor vaccines, but they are expensive, which limits their use

Method used

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  • Method of establishing animal model of HER2 positive tumor by using normal mouse
  • Method of establishing animal model of HER2 positive tumor by using normal mouse
  • Method of establishing animal model of HER2 positive tumor by using normal mouse

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Screening of human-derived HER2-positive tumor cells

[0044] 10 HER2-positive human tumor cell lines were selected, including 7 breast cancer cell lines: SK-BR-3, MDA-MB-453, BT-474, HCC1008, HCC1569, HCC2218, HCC1419, and 1 ovarian cancer cell line: SKOV- 3. Two gastric cancer cell lines: SGC-7901, NCI-N87, all cells were purchased from ATCC.

[0045] Cell culture: The above cell lines were recovered from cryopreservation tubes, cultured in RPMI 1640 medium, and subcultured until the cells were in a good growth state for later use.

[0046] Inoculation: 60 8-week-old Balb / c mice were randomly divided into 10 groups with 6 mice in each group. After one week of adaptation, the above 10 kinds of tumor cells were inoculated respectively. The inoculated cell concentration was 5×10 6 1 / ml, 0.1ml was inoculated into each mouse, and the inoculation site was the armpit of the left forelimb.

[0047] After tumor inoculation, the tumor growth was observed and record...

Embodiment 2

[0049] Example 2 Optimization of the inoculum amount of human gastric cancer cell NCI-N87 transplanted into mice

[0050] In order to establish the best NCI-N87 animal model, the inoculation amount of human gastric cancer cell NCI-N87 was optimized.

[0051] Twenty-four 8-week-old Balb / c mice were randomly divided into 4 groups, 6 mice in each group. After one week of adaptation, the inoculation concentration was 1.25×10 6 pcs / ml, 2.5×10 6 pcs / ml, 5×10 6 pcs / ml, 1×10 7 0.1 ml of human gastric cancer cell NCI-N87 cells / ml was inoculated at the axilla of the left forelimb. After tumor inoculation, the tumor growth was observed and recorded daily, and 2 weeks after inoculation, all animals were sacrificed to end the experiment, and the test results were evaluated.

[0052] Result: 1.25×10 6 Three animals in the unit / ml group developed tumors, and the tumor formation rate was 50%. 2.5×10 6 pcs / ml, 5×10 6 pcs / ml, 1×10 7 All the animals in the group / ml group have tumor grow...

Embodiment 3

[0053] Example 3 Solid Phase Synthesis of Tumor Vaccine Peptide E75

[0054] E75 entrusts Shanghai Aobo Biotechnology Co., Ltd. to synthesize it by conventional solid-phase technology, and the purity of the synthesized peptide is >98%, see figure 1 .

[0055] Evaluation of NCI-N87 animal model for therapeutic effect of tumor vaccine

[0056] Inoculation: 40 8-week-old female Balb / c mice were used in this study. Human gastric cancer cells NCI-N87 in the logarithmic growth phase were collected and prepared into 5×10 6 0.1ml of the cell suspension was planted in the left armpit of the mice; the tumor growth was observed every day, and the immunotherapy was started when the tumor diameter grew to about 8mm.

[0057] Immunization: 40 mice were randomly divided into 4 groups, 10 in each group, respectively negative control group (PBS) and E75 low, medium and high dose groups, E75 respectively with 100, 300, 1000 μg / ml and Freund's incomplete Adjuvant (FIA) was mixed and emulsifi...

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Abstract

The invention discloses a method of establishing animal model of an HER2 positive tumor by using a normal mouse. The method comprises the following steps: (1) cell cultivation: cultivating a human gastric carcinoma cell NCI-N87 in a cell culture medium and collecting cells in a logarithmic phase; (2) inoculation: inoculating the human gastric carcinoma cell NCI-N87 to the normal mouse; (3) evaluation: after inoculation for two weeks, the diameter of the tumor is greater than or equal to 0.6cm and the tumor formation rate is greater than or equal to 80%, showing that the animal model of the HER2 positive tumor is successively established, wherein an immunosuppressive step of the normal mouse is not included. The method of directly inoculating the normal mouse to prepare the animal model by means of the NCI-N87 is simple to operate and stable and reliable, the test cost can be greatly lowered, and many limitations of current other tumor vaccine animal models are overcome. The method of detecting the treatment potency or preventing potency of the HER2 positive tumor vaccine can be used for screening tumor vaccine candidates in a lab and has a wide application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for establishing an animal model of HER2-positive tumors using normal mice. Background technique [0002] Malignant tumors are one of the major diseases that threaten human life and health. Traditional cancer treatment methods such as surgery, radiation therapy, and drug therapy have certain limitations. Due to poor targeting, radiation therapy and drug therapy can easily damage normal human cells and cause adverse reactions. With the in-depth study of the molecular mechanism of tumor occurrence and development and the rapid development of biotechnology, biological therapy has become the fourth mode of tumor treatment. Polypeptide tumor vaccines are vaccines composed of polypeptides derived from tumor cell-specific antigens, virus-associated antigens, oncogenes or tumor suppressor gene mutant proteins. With the assistance of cytokines, chemokines and other adjuvants, they c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/13A61K49/00
CPCA61K35/13A61K49/0008
Inventor 宋燕许元生卢肖宇
Owner LINK HEALTH GRP
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