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Constructing method and application of zebra fish NK/TCL tumor model

A construction method, zebrafish technology, applied in the field of medical biology, to achieve the effect of short experiment cycle, less tumor cell consumption and high repeatability

Active Publication Date: 2018-11-20
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the construction of zebrafish patient-derived xenograft model (zPDX model) and related molecular biology research are still in their infancy

Method used

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  • Constructing method and application of zebra fish NK/TCL tumor model
  • Constructing method and application of zebra fish NK/TCL tumor model
  • Constructing method and application of zebra fish NK/TCL tumor model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Construction of zebrafish NK / TCL tumor model

[0034] 1. Materials and Methods

[0035] 1.1 Reagents

[0036] Bambanker was purchased from WAKO Company for cryopreservation of tumor tissues.

[0037] Liberase was purchased from Sigma and used for tumor tissue digestion.

[0038] Dil was purchased from Invitrogen and used for fluorescent staining of tumor cells.

[0039] 1.2 Tumor tissue digestion and tumor cell staining

[0040] Fresh NK / T-cell lymphoma tissue after surgery or biopsy was cut into 1mm*1mm size, placed in Bambanker solution, and frozen (stored in a -80°C refrigerator).

[0041] The NK / T cell lymphoma tissue used in the experiment was digested with Liberase into a single cell suspension and counted, resuspended in PBS at a density of 1*10^7 / mL, stained with Dil dye, and NK / TCL cells were detected under a fluorescence microscope. Observe red fluorescence.

[0042] 1.3 Microinjection of human tumor cells

[0043] Take 48-hour zebrafish embry...

Embodiment 2

[0064]Example 2 Application of zebrafish NK / TCL tumor model in drug screening

[0065] The method of Example 1 was used to establish the zebrafish NK / TCL tumor model, and after 24 hours of modeling, the tumor formation was confirmed by fluorescence microscope observation, and the modeling was confirmed to be successful. 45 items were selected, randomly divided into groups, and treated with anti-lymphoma chemotherapy drug Doxorubicin (Doxorubicin).

[0066] 1. Grouping and medication

[0067] Control group: 20 animals were directly cultured with egg water without drugs, and the volume of the culture medium was 4 mL.

[0068] Pharmacotherapeutic group:

[0069] Doxorubicin (Doxorubicin) was dissolved in egg water to form drug solutions with concentrations of 0.02ng / mL and 0.04ng / mL, respectively;

[0070] The first treatment group, 10, was cultured with drug-containing egg water, the administration concentration was 0.02ng / mL, and the volume of culture solution was 2mL;

[0...

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Abstract

The invention belongs to the technical field of medical biology and particularly relates to a constructing method and application of a zebra fish NK / TCL tumor model. Through research, the invention successfully constructs an NK / TCL human tumor xenotransplantation model by taking zebra fish as a mode organism for the first time. The construction method has the advantages of simple and feasible experiment operation, short experiment period, extremely low consumption of tumor cells and high repeatability, and can be widely applied to the fields of molecular biology study, targeted drug screeningand the like.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and in particular relates to a construction method and application of a zebrafish NK / TCL tumor model. Background technique [0002] NK / T cell lymphoma (natural killer / T-cell lymphoma, NK / TCL) is a kind of CD56-positive / cytoplasmic CD3-positive lymphocyte malignant proliferation disease, which occurs in Asia, and Epstein-Barr virus (EB Virus) infection is closely related. In the past two decades, a series of molecular biology studies have reported a variety of genetic and molecular biological abnormalities associated with the progression of NK / TCL disease. Comparative genomics studies (Comparative genomichybridization, CGH) have reported a series of disease-associated DNA gene copy number alterations (copy number alterations), including 2q5, 13q4, 10q3, 21q2, 3q2, 5q2 and 17q2 expansion of chromosome segments, as well as 1p4, Deletions of chromosomal segments such as 17p4, 12q3, 13q2, and 6q...

Claims

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Application Information

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IPC IPC(8): C12N15/873A01K67/027A61K49/00
CPCA61K49/0008A01K67/0271C12N15/873A01K2267/03A01K2207/12A01K2227/40
Inventor 赵维莅熊杰孙晓建程澍
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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