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Application of NO donor compound in preparation of drugs for inhibiting invasion and metastasis capabilities of tumor cells rich in sulfhydryl molecules

A technology of tumor cells and metastatic ability, applied in the direction of anti-tumor drugs, drug combinations, tripeptide components, etc., can solve the problem of not changing the state of sulfhydryl group

Active Publication Date: 2018-03-23
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Currently, there is no technology aimed at altering the sulfhydryl status of proteins rich in sulfhydryl molecules to inhibit tumor cell invasion and metastasis

Method used

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  • Application of NO donor compound in preparation of drugs for inhibiting invasion and metastasis capabilities of tumor cells rich in sulfhydryl molecules
  • Application of NO donor compound in preparation of drugs for inhibiting invasion and metastasis capabilities of tumor cells rich in sulfhydryl molecules
  • Application of NO donor compound in preparation of drugs for inhibiting invasion and metastasis capabilities of tumor cells rich in sulfhydryl molecules

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Take the protein molecule "CYR61" rich in sulfhydryl groups as an example: the sulfhydryl nitrosylation reaction of CYR61 protein: MDA-MB-231 cells were washed by adding sulfhydryl nitrosoglutathione (S-nitrosoglutathione, GSNO) to make it The final concentration was 30 μM, and incubated at 37° C. in the dark for 30 minutes. Add 200 μL of HENS buffer solution to the cell suspension, place the centrifuge tube in a small box with crushed ice, and ultrasonically disrupt for 30 min in a cell disruptor. Centrifuge at 10,000 g for 10 min at 4°C, collect the supernatant in a centrifuge tube, and measure the protein concentration by the BCA method. Add 2 μL of 1 M MMTS to the protein sample, vortex for 1 min to fully mix with the protein, and incubate at room temperature for 30 min in the dark. Add 600 μL of pre-cooled acetone, and place in a -20°C refrigerator for 1 h in the dark. Centrifuge at 10,000 g for 10 min at 4°C, pour off the acetone, and dry in the air. Resuspend ...

Embodiment 2

[0041] The effect of CYR61 on the adhesion of breast cancer cell MDA-MB-231 to the cell matrix, and the effect of thiol-rich protein molecules (such as CYR61) on breast cancer cell MDA-MB-231 by thiol-nitrosoglutathione sulfhydryl nitrosylation. This effect reduces the adhesion of cancer cells to the cell matrix.

[0042] Spread gelatin: Take a 24-well plate, add 200 μL gelatin to each well, shake well, and distribute evenly in the wells. Incubate overnight at 37°C in an incubator. Blot the excess gelatin, add 200 μL of PBS containing 2% BSA to each well, and place in an incubator at 37°C for 1 hour to block. Aspirate the liquid and wash three times with PBS. Cell count: MDA-MB-231, OE-CYR61, MDA-MB-231, Si-CYR61 MDA-MB-231 cells were washed and digested, then resuspended twice in PBS. Pipette 2 parts of 12 μL cells into new EP tubes, add 12 μL reagent T to one part, and add 12 μL reagent NC to one part, mix well, and add to the sample well of the cell counting plate. Ins...

Embodiment 3

[0045] CYR61 affects the adhesion of breast cancer cell MDA-MB-231 to endothelial cells, and S-nitrosoglutathione (S-nitrosoglutathione) has an effect on the sulfhydryl of breast cancer cell MDA-MB-231 rich in sulfhydryl molecules (such as CYR61). Nitrosylation modification inhibits the adhesion of tumor cell MDA-MB-231 to endothelial cells.

[0046] Endothelial cell plating: take logarithmically grown umbilical vein endothelial cells, wash and digest them with PBS, spread the cells in a 24-well plate, and wait until they are covered with a layer of endothelial cells. The cells were washed three times with PBS, ECM medium containing 10 μg / mL TNF-α was added, and incubated in a 37°C incubator for 4 h to activate endothelial cells. MDA-MB-231, OE-CYR61 MDA-MB-231, Si-CYR61 MDA-MB-231 cells were stained with rhodamine b, plated and observed. The umbilical vein endothelial cells with logarithmic growth were inoculated in a 24-well plate covered with gelatin. After the endothelial...

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Abstract

The invention relates to application of an NO donor compound in preparation of drugs for inhibiting invasion and metastasis capabilities of tumor cells rich in sulfhydryl molecules. The NO donor compound is prepared from one or more of sulfhydryl-nitroso glutathione, nitric oxide, sodium nitrite, nitroglycerin, the sulfhydryl-nitroso glutathione, sulfhydryl-nitroso cysteine, sulfhydryl-nitroso captopril, sulfhydryl-nitroso-N-acetylpenicillamine, monosaccharide-sulfhydryl-nitroso-N-acetylpenicillamine and a conjugate of the monosaccharide-sulfhydryl-nitroso-N-acetylpenicillamine. The NO donor compound and the sulfydryl molecules in the tumor cells undergo sulfhydryl-nitrosylation reaction, so that sulfydryl is transformed into sulfhydryl nitroso, and further the invasion and metastasis capabilities of the tumor cells rich in the sulfhydryl molecules can be inhibited. The method for inhibiting the invasion and metastasis capabilities of the tumor cells rich in the sulfhydryl molecules, provided by the invention, provides a new direction for treating tumor metastasis and has obvious social benefits.

Description

technical field [0001] The invention relates to the application of a NO donor compound in the preparation of drugs for inhibiting the invasion and metastasis of tumor cells rich in thiol molecules. Background technique [0002] Tumor metastasis is an important biological feature of malignant tumors. Most cancer patients die not from primary cancer but from metastatic cancer. The occurrence of tumor metastasis involves complex and subtle changes in tumor cells and their microenvironment, which lead to tumor metastasis, invasion and survival in the blood / lymphatic circulatory system, as well as the growth process at the metastatic target site . Only a small fraction of cells in the primary tumor have the ability to metastasize. In animal models, only 0.01% or even less cells can enter the circulation for metastatic tumor cells. Endogenous genomic instability increases the likelihood of acquiring metastatic ability. Tumor cells with unstable and heterogeneous genomes have f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K38/06A61P35/00A61P35/04
CPCA61K38/06A61K45/00
Inventor 贾力余素红贺苏丹李书慧
Owner FUZHOU UNIV
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