Application of idebenone in preparation of medicines for inhibiting tumor cell metastasis and spread
A technology of tumor cells and idebenone, which is applied in the application field of drugs, can solve the problems that the pharmacological effects of idebenone have not been discovered, and achieve the effect of reducing the research and development cycle and reducing the amount of death
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Embodiment 1
[0025] Example 1: Application of idebenone in the preparation of a drug for inhibiting the metastasis and spread of colon cancer cells. The specific experimental plan is as follows:
[0026] 1. Experimental materials
[0027] Colon cancer cells (HT29) were used in the experimental stage of the present invention, and the drug used in the positive control was nocodazole (experimental concentration: 10uM). The negative control group received no treatment.
[0028] The experiment used idebenone compound C 19 h 30 o 5 , CAS NO. is 58186-27-9, the concentrations are 100uM, 80uM, 60uM, 50uM, 4uM, 20uM, 10uM.
[0029] 2. Experimental method
[0030] For colon cancer cells cultured for 24 hours (cell density: 10 5 ) for drug stimulation for 2 hours:
[0031] (1) Centrifuge the colon cancer cells in a better state in the culture dish, and add an appropriate amount of culture medium to make a cell suspension;
[0032] (2) Get the cell suspension count, according to 10 5 The den...
Embodiment 2
[0040] Example 2: Transwell cell migration assay was used to determine the effect of idebenone on the migration ability of colon cancer cells.
[0041] Will 1×10 5 The HT29 cells were inoculated in Transwell chambers with a 12.0 μm membrane at the bottom, and continued to be cultured. After the cells adhere to the wall, add DMEM complete medium with different concentrations of idebenone to the upper chamber. To maintain osmotic pressure, add 0.05%-0.2% BSA, and add DMEM complete medium containing 15% FBS to the lower chamber and continue culturing for 24 hours . The unmigrated cells on the upper surface of the membrane were carefully wiped off with a cotton swab, and the cells that migrated to the lower surface of the membrane were fixed with 2% PFA at 37°C for 10 minutes and then stained with Giemsa. Ten visual fields were randomly selected under a high-power microscope to count the migrated cells.
[0042] Cell migration rate = [the average number of cells in the experime...
Embodiment 3
[0046] Example 3: The damage repair experiment detects the effect of idebenone on the migration ability of tumor cells.
[0047] Take HT29 cells in the logarithmic growth phase, and add 1×10 cells to each well of a 24-well plate 5 After the cells grow into a single layer, draw a "one"-shaped scratch, wash with PBS, add different concentrations of idebenone to the control group and the experimental group, and take pictures. After 22 hours of culture, the same part is taken again to measure the migration distance. The experiment was repeated 3 times, and the cell migration rate=[(0h average value of the experimental group-22h average value of the experimental group) / (0h average value of the blank control group-22h average value of the blank control group)]×100%.
[0048]Idebenone inhibited the migration of LTEP and A549 cells at different concentrations (10-100 μg / ml) for 22 hours, and the results showed that idebenone had a dose-dependent inhibitory effect on colon cancer cell ...
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