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Nano-silver combined autophagy inhibitor capable of killing and damaging tumor

An autophagy inhibitor, nano-silver technology, used in anti-tumor drugs, drug combinations, medical preparations containing active ingredients, etc.

Active Publication Date: 2015-01-07
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether silver nanoparticles can induce autophagy, and how the induced autophagy plays a role in the survival of cells has not been reported.
In addition, there is no report on the role of autophagy in the process of killing tumor cells by silver nanoparticles.

Method used

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  • Nano-silver combined autophagy inhibitor capable of killing and damaging tumor
  • Nano-silver combined autophagy inhibitor capable of killing and damaging tumor
  • Nano-silver combined autophagy inhibitor capable of killing and damaging tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Embodiment 1: Electrochemical synthesis of water-soluble nano-silver particles

[0102] We synthesized PVP-wrapped silver nanoparticles by an electrochemical method based on continuous flow technology, and scanned electron microscopy ( figure 1 ) Analysis shows that the particle size of our synthesized nano-silver is 26.5 ± 8.4nm. Dynamic Light Scattering Analysis ( figure 2 ) shows that the hydrated particle size of nano-silver is 58.8±1.7nm, and the polydispersity index is 0.29±0.04. The zeta potential of nano silver in water is -12.9±1.1mV.

Embodiment 2

[0103] Example 2: Nano silver treatment causes intracellular autophagy

[0104] We constructed a HeLa cell line stably transfected with EGFP-LC3. When autophagy occurs, you can see the accumulation of green fluorescent spots in the cells ( Figure 3a ). In a 24-well plate, when the EGFP-LC3 / HeLa cells grow to about 60% density, add 10 μg / ml nano-silver to treat the cells, and take pictures at 0, 2, 4, 6, 8, 10, and 12 hours, respectively. And statistical analysis was performed on the ratio of cells containing fluorescent dots, we found that with the increase of time, the proportion of cells containing fluorescent dots was larger, indicating that the induced cell autophagy was stronger ( Figure 3b ). In a 24-well plate, when the HeLa cells grow to about 60% density, add different concentrations (0, 5, 10, 15, 20 μg / ml) of nano-silver to treat the cells for 24 hours, and then use the cell lysate to lyse the cells. Similarly, by western blotting detection, we found that with ...

Embodiment 3

[0105] Example 3: Wortmannin inhibits autophagy and promotes the killing of cells by nano-silver

[0106] In 24-well plates, when the HeLa cell density grew to about 60%, we used 1 μM wortmannin (purchased from Sigma-Aldrich), 10 μg / ml nanosilver, 1 μM wortmannin plus 10 μg / ml nano The cells were co-treated with silver for 24 hours, and then the apoptosis of the cells was detected by three methods: MTT, Annexin A5 (ANXA5)-FITC PI flow cytometry analysis, and HO / PI staining analysis. MTT results showed that the co-treatment of 1 μM wortmannin and 10 μg / ml nano-silver was compared with single-treatment with nano-silver, and the cell viability was weakened by 20.91%, and wortmannin single treatment had no effect on cell viability ( Figure 4a ). ANXA5-FITC PI flow analysis ( Figure 4b ), consistent with the results of MTT analysis, wortmannin will reduce the cell death caused by silver nanoparticles (the relative proportion of cells located in the lower right and upper right a...

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Abstract

The invention relates to a nano-silver combined autophagy inhibitor capable of killing and damaging tumor, and particularly relates to applications of a nano-silver combined autophagy inhibitor and / or a reagent for knocking down / knocking out autophagy related gene expression on preparation of an anti-tumor medicament, and applications of the autophagy inhibitor and / or the reagent for knocking down / knocking out autophagy related gene expression on promotion of nano silver to kill and damage tumor cells.

Description

technical field [0001] The invention relates to the use of nano-silver and an autophagy inhibitor or a reagent for knocking down / knocking out the gene expression necessary for autophagy for the preparation of antitumor drugs. Background technique [0002] Nano-silver is a simple substance of metallic silver with a particle size of nanometer scale. Due to its good bactericidal effect, nano-silver is used in the medical field, such as antibacterial medicine and medical equipment, antibacterial plastic and rubber products, antibacterial textiles, clothing, footwear, etc. Nano silver also has many other biological effects, such as antifungal, antiviral, anti-inflammatory. As one of the few nanomaterials that has been applied clinically, recent experiments on cells and mice have shown that nanosilver may also be a potential antitumor drug. Nanosilver has been proven to have a strong cell killing effect in a variety of cancer cells, including oxidative stress, DNA damage, cell c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K48/00A61P35/00A61K33/38
Inventor 温龙平林俊顾宁张云娇周伟金佩佩
Owner UNIV OF SCI & TECH OF CHINA
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