Anti-tumor metastasis medicament high flux screening model

An anti-tumor metastasis, high-throughput technology, applied in the direction of measurement devices, biochemical equipment and methods, microbial determination/testing, etc., can solve the problems of screening anti-tumor metastasis drugs, etc., and achieve the effect of high sensitivity and easy operation

Inactive Publication Date: 2009-03-04
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Studies have found that TF is related to tumor formation, invasion and metastasis, but there is no report on the use of cell-level tissue factor high-throughput drug screening models to screen anti-metastatic drugs

Method used

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  • Anti-tumor metastasis medicament high flux screening model
  • Anti-tumor metastasis medicament high flux screening model
  • Anti-tumor metastasis medicament high flux screening model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Materials and methods:

[0032] 1. Instrument

[0033]

[0034] 2. Materials and reagents:

[0035] MDA-MB-435 (human breast cancer cells, cell bank of the Basic Institute of Chinese Academy of Medical Sciences), IM-3 (human liver cancer cells, Zhongshan Hospital Liver Cancer Institute), LLC cells (fluorescence-labeled Lewis lung cancer cells, model animals of Nanjing University Center), SGC-7901 (human gastric cancer cells, cell bank of the Institute of Basic Medicine, Academy of Medical Sciences), BGC-823 (human gastric cancer cells, cell bank of the Institute of Basic Medicine, Academy of Medical Sciences), HL-60 (human leukemia cells, cell bank of the Institute of Basic Medicine, Academy of Medical Sciences) , A549 (human lung cancer cells, cell bank of the Basic Institute of Medical Sciences), L-02 (human liver cells, cell bank of the Basic Institute of Medical Sciences), Bel-7402 cells (human liver cancer cells, cell bank of the Basic Institute of Medical Scie...

Embodiment 2

[0117] According to the method of Example 1, different compounds were screened using the optimum conditions, and the results are shown in Tables 2 and 3.

[0118] The optimum conditions are: the cells use MDA-MB-435, and the cell density when inoculated is 4×10 6 / ml; the concentration of the prothrombin complex is 0.008g / ml; the cell lysate is added to the human prothrombin complex solution and the incubation time is 15min. The chromogenic substrate is S-2222, the concentration of the chromogenic substrate is 0.5mM, CaCl 2 The optimum concentration is 100mmol / L, and the optimum concentration of EDTA is 50mmol / L.

[0119] Operation method: Well-growing MDA-MB-435 cells were divided into 4×10 6 Inoculate in a 96-well culture plate with 180 μl of cell fluid per well, culture until the cells are in good condition and about 80% adherent, absorb the medium, add 200 μl of fresh medium to the blank well and the control group, and add 180 μl to the administration well Fresh culture...

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Abstract

The invention belongs to the field of medicine and discloses an anti-tumor transfer medicine high throughput screening model. The screening method of the model comprises the following steps: an MDA-MB-435 cell is inoculated to a culture plate; a chemical compound to be screened is added into culture fluid and no inhibitor is added in; the chemical compound is cultured for an appropriate time of period and repeatedly melted, to obtain cell lysis solution; the cell lysis solution is added into human thrombinogen compound solution to be incubated together; a chromogenic substrate is added in; at 405nm, absorbency is determined and inhibitory rate is calculated; the chemical compound with the inhibitory rate higher than 20 percent is a positive compound. In the invention, the human breast cancer cell MDA-MB-435 is selected to build the anti-tumor transfer medicine screening model which is quick and stable, has high sensitivity and is convenient to be operated; the model is characterized by trace quantity and accuracy, and is applicable in screening anti-tumor transfer medicine with high throughput.

Description

technical field [0001] The invention belongs to the field of medicine and relates to a high-throughput screening model of anti-tumor metastasis medicine. Background technique [0002] Tissue factor (TF for short), that is, coagulation factor III, is a single-chain transmembrane glycoprotein, a member of the cytokine superfamily, and is the initiating factor of physiological hemostasis, coagulation process and various pathological thrombus formation. [1] . TF is both a cell surface receptor for coagulation factor VII (FVII) and a cofactor for FVII or FVII(a). When the blood vessel is damaged or under pathological conditions, the blood is exposed to the surface of cells expressing TF, and the receptor TF forms a complex with its ligand VII or VIIa, thereby simultaneously activating blood coagulation factors X and IX, and the activated FXa and FIXa are further activated Prothrombin (FII), FIIa turns fibrinogen into fibrin, thereby completing the coagulation cascade, leading t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N21/00
Inventor 袁胜涛孙立张陆勇余伯阳施伟寇俊萍
Owner CHINA PHARM UNIV
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