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Method for establishing extranodal nasal type NK/T-celllymphoma animal model

An animal model, lymphoma technology, applied in the field of biomedicine, can solve the problems of easy pollution, difficult transplantation, restrictions, etc., and achieve the effect of low success rate, accurate results and low cost

Inactive Publication Date: 2010-05-12
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since the tumor cells are cultured in vitro, leaving the in vivo environment, after long-term subculture, the interaction between cells in the body is eliminated, and the biological behavior and natural growth process of extranodal nasal NK / T cell lymphoma cannot be completely simulated. , will also lead to changes in the biological characteristics and genetic characteristics of tumor cells, thereby affecting the accuracy of experimental results and the reliability of conclusions
[0004] A method for establishing an animal model of human extranodal nasal NK / T cell lymphoma is disclosed in the document "Transplantation and Passaging of Human Nasal NK / T Cell Lymphoma in Nude Mice" (Chinese Journal of Pathology, April 2004) , but this method focuses on the universal method established by general tumor animal models, without targeted optimization of the characteristics of human extranodal nasal NK / T cell lymphoma, such as: concentration selection of double antibodies, passage The selection of the cycle, the selection of the cell concentration when the cryopreserved cells are revived and passaged, and the selection of the gene sequence label site, etc., therefore, there are still difficulties in transplantation, easy pollution, and unstable passage cycle (can only be passed to the 9th generation at most). Accurately identify deficiencies such as successful modeling, and the tumor model established by this method has not found metastases to the lungs, kidneys and other organs in advanced tumors, and no brain metastases have been observed
At the same time, this method does not involve the identification of the species of transplanted tumors, and it is not yet possible to accurately confirm the success of the modeling, so that this animal model cannot be directly used in the research of tumor gene therapy and drug screening, etc. also restricted

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1 uses the method of the present invention to establish the animal model of human extranodal nasal type NK / T cell lymphoma

[0034] 1. Collection and identification of human extranodal nasal NK / T cell lymphoma tissue samples

[0035] One case of human extranodal nasal NK / T-cell lymphoma was collected from West China Hospital of Sichuan University. After pathological examination, immunophenotype and EBER in situ hybridization detection, it was diagnosed as human extranodal nasal NK / T cell lymphoma. The patient received radiotherapy and chemotherapy (CHOP). After 2 courses of treatment, the local lesions disappeared and clinical remission was achieved. The patient went to the doctor again because of fever and stomach discomfort, and was diagnosed with secondary extranodal nasal NK / T cell lymphoma of the stomach. The patient underwent surgery because of progressive pyloric obstruction. The resected gastric tumor tissue was selected and planted under the skin ...

Embodiment 2

[0047] Example two extranodal nasal NK / T cell lymphoma cryopreserved cells resuscitation and passage experiment:

[0048] The frozen tumor cells of the 5th, 6th, 8th, 10th, 16th, 18th, 20th, 25th and 30th passage of the animal model established in Example 1 were taken. Thaw the above-mentioned frozen cells rapidly (within 30 seconds) in a 37-41°C water bath, and then wash the cells 2-3 times with 1640 medium added with penicillin and streptomycin in the ultra-clean workbench, and then adjust Cell concentration up to 10 7-8 / ml, take 0.5ml~1.0ml and inoculate subcutaneously in the forelimb armpits of healthy nude mice aged 3~4 weeks. Note that the passaging operation should be completed within 30 minutes.

[0049] The results showed that the success rate of resuscitation was 100%, and the resuscitation nude mouse transplanted tumor was similar to the growth cycle and tumor bearing situation of the original source tumor (see image 3 ). The classical method generally uses a ...

Embodiment 3

[0050] Example three Identification of human extranodal nasal NK / T cell lymphoma animal model

[0051] (1) Identification items

[0052] 1 Species Identification of Transplanted Tumors in Animal Models

[0053] 1.1 Human gene identification

[0054] Because the human and mouse genomes are highly homologous, the transplanted tumor tissue of the animal model comes from a male patient. The result of querying the gene bank found that the human male sex-determining gene sequence tag site (Sequence Tagged Sites, STS) SY14 has no homology with the mouse. Therefore, PCR technology was used to detect the presence or absence of SY14 gene in transplanted tumors. If the existence of human gene SY14 in transplanted tumors of animal models was proved, it was confirmed that the model was established successfully.

[0055] 1.2 Detection of human Y chromosome by fluorescence in situ hybridization (FISH)

[0056] The probe used in this study is the JAB-Y(DYZ1)(Yq12 SatelliteIII) DNA probe d...

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Abstract

The invention relates to a method for building the animal model of external nose shape NK / T lymphocyte cancer, which comprises: a, seeding, b, generating, c, checking, while it optimizes the parameters of each step, to overcome the defects of prestn technique. The invention has short generating time, with high on-time seeding success ratio. Compared with present technique, it can build the animalmodel of external nose shape NK / T lymphocyte cancer. And the model can replay the key biological character of external nose shape NK / T lymphocyte cancer, with stable growing period and wider application.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for establishing an animal model of human extranodal nasal type NK / T cell lymphoma. Background technique [0002] Human extranodal nasal-type NK / T cell lymphoma (human extranodal nasal-type NK / T cell lymphoma) is a tumor with unique morphology, immunophenotype and biological behavior, which mostly occurs in extranodal organs or tissues. Epstein-Barr virus (EBV) infection is highly related, but whether EBV is a pathogenic factor or an accompanying phenomenon of this lymphoma has not been clearly determined so far. The tumor has a high incidence in Asia, especially China, and is rare in Western Europe and North America. In the past 21 years, a total of 1012 cases of extranodal nasal NK / T cell lymphoma were treated in the pathology department of West China Hospital of Sichuan University, accounting for 13.83% of all lymphomas. This type of lymphoma is an aggressive ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/00
Inventor 刘卫平何妙侠唐琼兰杨帆王晓凌赵莎江炜李甘地
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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