A preparation method and application of an antineoplastic drug screening model targeting 4.1r and CD29

An anti-tumor drug and model technology, applied in the field of medicine, can solve the problems of high maintenance cost, unfavorable transportation and storage, etc., and achieve the effect of reducing production cost and wide applicability

Inactive Publication Date: 2011-12-07
ZHENGZHOU UNIV
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Problems solved by technology

However, the feeding and management of gene knockout mouse animal models requires special technical requirements and environmental facilities, and the daily maintenance cost is too high, which is not conducive to transportation and storage

Method used

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  • A preparation method and application of an antineoplastic drug screening model targeting 4.1r and CD29
  • A preparation method and application of an antineoplastic drug screening model targeting 4.1r and CD29
  • A preparation method and application of an antineoplastic drug screening model targeting 4.1r and CD29

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Embodiment Construction

[0030] The present invention is further described below in conjunction with embodiment (accompanying drawing):

[0031] 1. Experimental materials

[0032] Wild-type C57BL / 6J inbred mice (C57BL / 6J_4.1R+ / +), 4.1R knockout C57BL / 6J inbred mice (C57BL / 6J_4.1R- / -), both 2 months old SPF mice (special pathogen freemouses) were housed in a barrier system. The 865 cell line (ATCCNumber: CRL-7601) containing SV40 large T antigen was cultured in MDEM (Dulbecco's Modified Eagle's Medium) medium at 37°C, 5% CO 2 cultivated under conditions.

[0033] 2. Experimental method

[0034] 2.1 Primary culture of mouse embryonic fibroblasts (MEFs):

[0035] ① 2-month-old C57BL / 6J_4.1R- / - mice were mated in a male-female 1:1 pair cage at 6:00 p.m., and observed whether there were sperm plugs (milky white jelly) in the reproductive tract of the female mice every morning. The embryonic age recorded on the day was 0.5d;

[0036] ②When the mouse embryos develop to 13.5d-15.5d, anesthetize the preg...

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Abstract

A preparation method and application of an anti-tumor drug screening model targeting 4.1R and CD29, using protein 4.1R gene knockout mice as experimental materials to prepare C57BL/6J 4.1R-/-MEF primary cells, using Immortalized C57BL/6J 4.1R-/-MEF obtained by SV40 large T antigen transformation method. The purpose of the invention is to provide an in vitro cell screening model for new drug screening and gene therapy with protein 4.1R and CD29 as drug targets. The advantage is that it avoids the limitations of drug screening and production with 4.1R as the therapeutic target due to the high cost of raising gene knockout animals and the limited passage of primary cells in vitro. Reduce the production cost of targeted new drugs. In the C57BL/6J 4.1R-/-MEF established by the present invention, the activation of CD29 on the cell surface is inhibited due to the absence of 4.1R, which can be used to target CD29 for research and cell proliferation and migration, cytokine secretion, cell signaling Screening models for new anti-tumor drugs related to transduction regulation and tumor drug resistance.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to the preparation of an anti-tumor drug screening model, that is, the preparation of a C57BL / 6J_4.1R- / -MEF cell model, in order to serve as a cell screening model for the preparation of new drugs and gene therapy with 4.1R and CD29 as targets . Background technique [0002] Protein 4.1 (referring to the 4.1 color band after SDS-PAGE of erythrocyte membrane protein) is a basic component of the spectrin-based cell membrane skeleton in erythrocytes [1] , is a bridge connecting the erythrocyte transmembrane protein and the spectrin-actin skeleton network, and plays an important role in maintaining the shape, adhesion, fragility and normal physiological functions of erythrocytes [2] . Loss of human protein 4.1 expression leads to hemolytic anemia [3] . Because protein 4.1 was first discovered in mature red blood cells, it was named 4.1R. In recent years, three proteins with high homology to 4...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
Inventor 康巧珍安秀丽周晴晴汲振余阎红霞陈鲤翔王婷
Owner ZHENGZHOU UNIV
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