Application of pumpkin protein in preparing medicament for treating chronic myeloid leukemia
A technology for chronic myeloid and leukemia, which is applied in the application field of pumpkin protein in the preparation of drugs for the treatment of chronic myeloid leukemia, and can solve problems such as limited application and toxicity
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Embodiment 1
[0009] Example 1: Inhibitory effect of pumpkin protein combined with STI571 on K562 cells
[0010] Method: The K562 cells in the logarithmic growth phase were inoculated on a 96-well culture plate, and the inoculation amount per well was 6×10 3 / 120 μL. Experimental components CUS group (2 and 4 mg / L), STI571 group (0.05, 0.1, 0.2 and 0.4 μmol / L) and CUS+STI571 group (combination of CUS and STI571 at different concentrations above), add 20 μL to each well, set 4 duplicate wells; each well of the negative control group was added with the same amount of solvent, and 8 duplicate wells were set up, and 140 μL of nutrient solution was added to the blank control wells for zero adjustment of the instrument. Set 5% CO 2 , Incubate at 37°C for 72 hours, add 10 μL of 5 mg / mL MTT to each well, continue to cultivate for 4 hours, add 140 μL of lysate (10% SDS, 5% isobutanol, adjusted to pH 4.5 with HCL) to each well, and place at 37°C for 24 hours , Measure the absorbance of each well w...
Embodiment 2
[0015] Example 2: Effect of Pumpkin Protein Combined with STI571 Inducing K562 Cell Apoptosis
[0016] Methods: About 10 K562 cells treated with CUS (2 and 4 mg / L), STI571 (0.1 and 0.2 μmol / L) and CUS+STI571 (the combination of CUS and STI571 at different concentrations above) were collected for 72 hours, respectively. 6 , after washing with PBS, add pre-cooled (4°C) 70% ethanol, fix at -20°C for more than 12 hours, centrifuge to remove ethanol, resuspend after washing with PBS twice, and use PI solution (containing 0.005% PI, 0.1% TritonX- 100, 0.1mmol·L -1 EDTA and 0.01% RNase A) staining at room temperature in the dark for 30 min, flow cytometry to measure the percentage of apoptotic cells. Refer to the literature (Jin Zhengjun. Addition in combined drug use. Chinese Journal of Pharmacology, 1980; 1(2): 70-76) to calculate the q value and judge the effect of combined drug use: q=E a+b / (E a +E b -E a ×E b ), where E a+b is the percentage of apoptotic cells when the d...
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