Method for screening anti-tumor metastasis compounds

A technology for anti-tumor metastasis and compounds, which is applied in the field of screening anti-tumor metastasis compounds, and can solve the problems that cannot be used for screening

Inactive Publication Date: 2017-02-22
EXPLORING HEALTH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This old principle cannot be used to screen new drugs that inhibit tumor metastasis

Method used

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  • Method for screening anti-tumor metastasis compounds
  • Method for screening anti-tumor metastasis compounds
  • Method for screening anti-tumor metastasis compounds

Examples

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Embodiment 1

[0029] The present invention uses the highly metastatic cell line S18 to establish a spleen-liver metastasis nude mouse model (spleen-livermetastasis nude mouse model), and then gives the cells two anti-metastasis treatments (AM0097 and AM0098) designed by the inventors, about one One month later, the livers of all animals were taken out at the same time to calculate the number of metastatic nodules, such as figure 1 , 2 As shown, there was a statistically significant difference compared with the control, indicating that both treatments had anti-metastatic properties.

[0030] Cell immunofluorescence staining experiments showed that the expression of EMT markers in the S18 cells treated with two anti-metastatic treatments changed significantly in two-dimensional culture: the expression of the mesenchymal marker Vimentin was significantly reduced, and the expression of the epithelial marker E-cadherin was significantly reduced. Significantly increased (scale bar, 50μm), such a...

Embodiment 2

[0032] An embodiment of the method for screening compounds for anti-tumor metastasis of the present invention, the method comprises the following steps:

[0033] (1) Construct tumor cells capable of quantitatively displaying the EMT process, and measure the expression levels of EMT markers of the tumor cells, and the EMT markers include markers representing epithelial characteristics and markers representing mesenchymal; wherein the representative Epithelial markers (epithelial markers) are one or more combinations of E-Cadherin, Desmoplakin, Cytokeratins, Occludin and Claudin; the mesenchymal markers (mesenchymal markers) are Vimentin, N-cadherin, One or more combinations of Fibronectin, Snail1, Snail2, Twist1, Twist2, ZEB1, ZEB2, LAMC2, and Bmi-1, and the assay method is immunofluorescence assay or direct assay of fluorescein.

[0034] (2) co-incubating the compound to be tested with the tumor cells in the step (1) for 1-48 hours;

[0035] (3) After incubation, determine th...

Embodiment 3

[0038] An embodiment of the method for screening compounds for anti-tumor metastasis of the present invention, the method comprises the following steps:

[0039](1) Construct tumor cells capable of quantitatively displaying the EMT process, and measure the expression levels of EMT markers of the tumor cells, and the EMT markers include markers representing epithelial characteristics and markers representing mesenchymal; wherein the representative The epithelial markers (epithelial markers) are one or more combinations of E-Cadherin, Desmoplakin, Cytokeratins, Occludin and Claudin; the mesenchymal markers (mesenchymal markers) are Vimentin, N-cadherin , Fibronectin, Snail1, Snail2, Twist1, Twist2, ZEB1, ZEB2, LAMC2 and Bmi-1, the assay method is immunofluorescence assay.

[0040] (2) Plant the tumor cells in step (1) in a 24-well plate, a 96-well plate or a 384-well plate, add various compounds before or after cell attachment, and incubate together for 1-48 hours.

[0041] (3)...

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Abstract

The invention provides a method for screening anti-tumor metastasis compounds. By means of the change of the expression level of an EMT marker in tumor cells, a stable and effective system for screening the anti-tumor metastasis compounds is established, screening of a large quantity of compounds can be achieved, anti-metastasis new drug precursors can be found within a short period, and a technical solution is provided for developing a series of anti-cancer new drugs.

Description

technical field [0001] The invention relates to a method for screening compounds, in particular to a method for screening compounds against tumor metastasis. Background technique [0002] At present, most cancer patients die of distant metastasis. The anti-tumor drugs that have been developed in the market are all based on their inhibitory effect on tumor growth or tumor angiogenesis, but are often ineffective on tumor metastasis. Major pharmaceutical companies have failed to develop new drugs for inhibiting tumor metastasis. The main bottleneck is the lack of a high-throughput stable system that can be used to screen anti-tumor metastasis compounds. [0003] The basic principle of existing high-throughput screening of anticancer compounds is to detect IC50, that is, the concentration at which the proliferation of cancer cells is inhibited by 50%. The lower the value of IC50, the stronger the inhibitory effect of the drug on tumor growth. This old principle cannot be used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/53
CPCG01N33/5011G01N33/53
Inventor 钱朝南
Owner EXPLORING HEALTH LLC
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