Applications of niclosamide for preventing pulmonary metastasis of malignant melanoma
A technology for melanoma and niclosamide, which is applied in the field of medicine to achieve the effect of reducing lung weight and lung coefficient, and reducing lung metastatic nodules
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Embodiment 1
[0023] Niclosamide inhibits the migration of melanoma cell A375.
[0024] Scratch test was used to determine the inhibitory effect of niclosamide on melanoma cell migration based on the experimental model of cell wound healing in vitro. The melanoma cells A375 in the logarithmic growth phase were taken, digested with trypsin and centrifuged to prepare a uniform cell suspension and inoculated in a 6-well plate, and incubated in a 37°C incubator for 24 hours to reach 80% confluence. Use a 200 μL sterile gun tip to quickly and clearly scratch the central area of the cells, wash with the corresponding fresh medium three times, and remove the scratched dead cells. Add 2 mL of medium containing different concentrations of niclosamide to the scratched wells. After 24 hours, the cells were observed and photographed under an inverted microscope, and the migrated cells were quantitatively analyzed. The number of cell migration in the negative control treated with 0.1% DMSO was defin...
Embodiment 2
[0029] Niclosamide inhibits the migration of melanoma cell B16-F10
[0030] Scratch test was used to determine the inhibitory effect of niclosamide on melanoma cell migration based on the experimental model of cell wound healing in vitro. The melanoma cells B16-F10 in the logarithmic growth phase were taken, digested with trypsin and centrifuged to prepare a uniform cell suspension and inoculated in a 6-well plate, and incubated in a 37°C incubator for 24 hours to reach 80% confluence. Use a 200 μL sterile gun tip to quickly and clearly scratch the central area of the cells, wash with the corresponding fresh medium three times, and remove the scratched dead cells. Add 2 mL of medium containing different concentrations of niclosamide to the scratched wells. After 24 hours, the cells were observed and photographed under an inverted microscope, and the migrated cells were quantitatively analyzed. The number of cell migration in the negative control treated with 0.1% DMSO was ...
Embodiment 3
[0035] Niclosamide inhibits the migration of melanoma cells A375Transwell
[0036] ==Using Transwell migration test method. The main material is Transwell chamber, which looks like a small cup that can be placed in the orifice plate. The bottom layer is a permeable membrane, usually polycarbonate membrane.
[0037] In this experiment, a Transwell chamber with a pore size of 8 μm was used. The Transwell chamber was placed on a 24-well plate, and 100 μL of serum-free medium containing 5×10 4 melanoma cells B16-F10 and different concentrations of niclosamide, and the negative control was 0.1% DMSO. Add 600 μL of complete medium containing 10% FBS and different concentrations of niclosamide to the lower layer of the small chamber. After culturing in the incubator for 20 h, the cells migrating to the back of the microporous membrane were fixed with 75% ethanol for 10 min, and then stained with 0.5% crystal violet·ethanol solution for 10 min. The stained microporous membrane was...
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