Application of flunarizine hydrochloride in preparation of medicines for inhibiting tumor cell metastases and spread
A technology of flunarizine hydrochloride and tumor cells, which is applied in the application field of medicine, can solve the problems such as the undiscovered pharmacological effects of flunarizine hydrochloride, achieve a large blocking effect, reduce the research and development cycle, and reduce the effect of death
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Embodiment 1
[0030] Example 1: Application of flunarizine hydrochloride in the preparation of drugs for inhibiting the metastasis and spread of colon cancer cells. The specific experimental plan is as follows:
[0031] 1. Experimental materials
[0032] Colon cancer cells (HT29) were used in the experimental phase of the present invention, and nocodazole (experimental concentration: 10uM) was used as the positive control. The negative control group received no treatment.
[0033] Wherein the experiment adopts flunarizine hydrochloride compound C 26 h 28 C l2F2 N 2 , CAS NO. is 30484-77-6, 52468-60-7 [flunarazine], the concentrations are 100 uM, 50 uM, 10uM, 5uM, 1uM, 0.5uM respectively.
[0034] 2. Experimental method
[0035] For colon cancer cells cultured for 24 hours (cell density: 10 5 ) for drug stimulation for 2 hours:
[0036] (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 suspen...
Embodiment 2
[0045] Example 2: Transwell cell migration assay was used to determine the effect of flunarizine hydrochloride on the migration ability of colon cancer cells.
[0046] 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 flunarizine hydrochloride 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 to cultivate 24h. 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 37oC for 10 minutes and then stained with Giemsa. Ten visual fields were randomly selected under a high-power microscope to count the migrated cells.
[0047] Cell migration rate = [the average numb...
Embodiment 3
[0051] Example 3: Damage repair assay was used to detect the effect of flunarizine hydrochloride on the migration ability of tumor cells.
[0052] 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 monolayer, draw a "one"-shaped scratch, wash with PBS, add different concentrations of flunarizine hydrochloride 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, cell migration rate = [(0h average value of experimental group - 22h average value of experimental group) / (0h average value of blank control group - 22h average value of blank control group)] × 100%.
[0053] Flunarizine hydrochloride inhibited the migration of LTEP and A549 cells at different concentrations (0.5-100μg / ml) for 22 hours, and the results showed that flunarizine hydrochloride had a d...
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