Application of doramectin in treatment of gliomas
A technology of doramectin and glioma, applied in the application field of doramectin in the treatment of glioma, can solve the problems that the effect of doramectin has not been reported, achieve good treatment and prognosis, and inhibit proliferation and migration, promote the effect of autophagy
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Embodiment 1
[0024] Example 1: Verification that doramectin inhibits the growth of glioma cells U87 and C6 cells in vitro.
[0025] 1. Experimental method:
[0026] U87 and C6 cells in the logarithmic growth phase were treated with 3.0×10 3 The number of cells was seeded in a 96-well plate and placed in 37°C, 5% CO 2 Cultivate in the incubator for 24h;
[0027] Discard the old medium in the 96-well plate, wash the cells twice with PBS, and treat the cells with doramectin or chloroquine;
[0028] Place the 96-well plate in 37°C, 5% CO 2 Cultivate in a constant temperature incubator, take out the 96 plates that have been cultured for different times, suck out the liquid in the wells, and wash them twice with PBS;
[0029] Take out the 96-well plate, discard the MTT solution, and add 150 μL of DMSO solution to each well;
[0030] Put the 96-well plate in a shaker in the dark, and shake it for 30 minutes;
[0031] Detect the absorbance value of the microplate reader at a wavelength of 49...
Embodiment 2
[0036] Example 2: Verification that doramectin inhibits the migration of U87 and C6 cells.
[0037] 1. Experimental method:
[0038] U87 and C6 cells in the logarithmic growth phase were treated with 2.0×10 5 The number of cells was seeded in a 6-well plate and placed at 37°C, 5% CO 2 Cultivated in an incubator;
[0039] After the cell density reaches 80%, discard the culture medium, wash twice with PBS, draw a scratch of about 1 cm from top to bottom in the center of each well with a pipette tip, wash twice with PBS again, and put into an inverted microscope to take pictures;
[0040] After discarding the PBS, treat U87 and C6 cells with different concentrations of doramectin, and culture them in the incubator for 24 hours;
[0041] After the supernatant was discarded, washed twice with PBS, put into an inverted microscope to take pictures again, the experiment was repeated three times.
[0042] 2. Experimental results:
[0043] The results showed that the scratch dista...
Embodiment 3
[0046] Example 3: Verification of doramectin-induced apoptosis in U87 and C6 cells.
[0047] 1. Experimental method:
[0048] U87 and C6 cells in the logarithmic growth phase were treated with 1.0×10 5 The number of cells was seeded in a 6-well plate and placed in 37°C, 5% CO 2 Cultivate in the incubator for 24h;
[0049] After the cells were treated with different concentrations of doramectin or chloroquine for 48 hours, they were washed twice with pre-cooled PBS;
[0050] Add 0.5 mL of DAPI dye to the 6-well plate, and incubate at 37°C for 15 minutes in the dark;
[0051] The 6-well plate was washed 3 times with PBS for 10 min each time, detected and photographed with a fluorescence microscope, and the experiment was repeated three times.
[0052] 2. Experimental results:
[0053] The results showed that after the cells were treated with doramectin, obvious changes such as nuclear condensation and DNA fragmentation appeared in the cells. The apoptosis rate of U87 cells...
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