Use of L-stepherine for inhibiting or treating metastatic breast cancer
A technology of levothrinine and metastatic breast cancer, which is applied in the directions of medical preparations containing active ingredients, organic active ingredients, pharmaceutical formulas, etc., to achieve good drug safety and the effect of inhibiting lung metastasis
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Embodiment 1
[0023] Effects of SPD on Cell Survival of Metastatic Breast Cancer in Vitro
[0024] This example studies the effect of SPD on the survival of human metastatic breast cancer MDA-MB-231 cells and mouse metastatic breast cancer 4T1 cells in vitro.
[0025] Cells in logarithmic growth phase were taken, digested with 0.25% trypsin-0.53mmol / L EDTA solution, and counted. MDA-MB-231 cells were inoculated in 96-well cell culture plates at 5000 / well (3000 / well for 4T1). After culturing for 24 hours, administration was carried out. The final concentrations of the SPD administration groups were 0.39, 0.78, 1.56, 3.125, 6.25, 12.5, 25, and 50 μmol / L. Six replicates were set up in each group, and the incubation was continued for 48 hours. After incubation, discard the medium, add 100 μL of pre-cooled 10% trichloroacetic acid (TCA, w / v) solution to each well, and fix at 4 °C for 1 h; discard the TCA solution, rinse with tap water 5 times, and dry naturally; Add 100 μL of 0.4% sulforhoda...
Embodiment 2
[0030] The effect of SPD on the migration ability of metastatic breast cancer cells in vitro
[0031] This example studies the inhibitory effect of SPD on the migration of human metastatic breast cancer MDA-MB-231 cells and mouse metastatic breast cancer 4T1 cells.
[0032] MDA-MB-231 or 4T1 metastatic breast cancer cells with normal morphology and in good condition were digested, centrifuged, resuspended and counted. According to the counting results, the cell suspension was diluted to 1 million cells / mL with culture medium, and added to a 6-well cell culture plate at a volume of 2 mL per well to make 2 million cells per well. The seeded 6-well plate was placed in a cell culture incubator for 12 hours to allow the cells to adhere to the wall. Use a 200 μL pipette tip to evenly draw a line of uniform thickness in the middle of the six-well plate covered with cells, so that the width of each well is consistent. Use 1 mL of PBS to wash the drawn cells three times, and add the...
Embodiment 3
[0038] The effect of SPD on cell invasion ability of metastatic breast cancer in vitro
[0039] This example studies the inhibitory effect of SPD on the invasion of human metastatic breast cancer MDA-MB-231 cells and mouse metastatic breast cancer 4T1 cells.
[0040] Dilute Matrigel 10 times with serum-free medium, take 50 μL of the diluted Matrigel and slowly add it to a 0.8 μM transwell small dish, evenly cover the bottom of the small dish, place the Matrigel-added small dish at 37°C for 1 hour to make Matrigel solidification.
[0041] MDA-MB-231 or 4T1 metastatic breast cancer cells with normal morphology and in good condition were digested, centrifuged, resuspended and counted. According to the counting results, dilute the cell suspension to 300,000 cells / mL with FBS-free medium, add it to a transwell dish at a volume of 200 μL per well, and add different amounts of SPD and SCH23390 standard solution respectively, so that the final concentration is SPD 12.5μM, SPD 25μM...
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