Application of indacaterol maleate to preparation of tumor resisting medicines
An anti-tumor drug, the technology of acid indene, applied in the field of medicine, can solve the problems that have not yet been reported on cancer cell research, and achieve the effect of inhibiting tumor growth
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Embodiment 1
[0026] Example 1 In vitro experiment: growth inhibitory effect of indacaterol maleate on colorectal cancer cells.
[0027] The detection is carried out by the WST-1 method, and the specific steps are as follows:
[0028] 1) Spread colorectal cancer cells HT29, RKO, DLD-1, and HCT116 into 96-well plates, with 3000 cells per well.
[0029] 2) Dissolve indacaterol maleate (purchased from Shanghai Taosu Biochemical Technology Co., Ltd.) in dimethyl sulfoxide (DMSO) to prepare a stock solution with a concentration of 10 mM, and then use RPMI1640 cell culture medium (containing 10% (v / v) FBS) to dilute the stock solution to prepare working solutions with final concentrations of 0 μM, 15 μM, 20 μM, 30 μM and 40 μM.
[0030] 3) After the cells in step (1) adhere to the wall, add the working solution of different concentrations prepared in step (2) into the corresponding wells in a volume of 100 μL per well. After the working solution is treated for 24, 48, and 72 hours, use WST respe...
Embodiment 2
[0033] Example 2 In vitro experiment: the cycle arrest effect of indacaterol maleate on colorectal cancer cells.
[0034] The steps are as follows: spread HT29 and RKO cells into 12-well plates, 2×10 per well 5 After the cells adhered to the wall, the cells were treated with a working solution of a certain concentration gradient (0 μM, 7.5 μM, 15 μM, 30 μM), and 1 mL of the working solution of the corresponding concentration was added to each well for 48 hours, and the cell cycle detection kit ( (purchased from Beyond Biotechnology Co., Ltd.) was used to detect cell cycle with C6Flow Cytometer flow cytometer (purchased from BD Accuri Company) after treating the cells.
[0035] The above experimental steps were biologically repeated three times. The sources of HT29 and RKO cells used in this step and the preparation method of the working solution were the same as in Example 1.
[0036] The results of the cell experiments figure 2 As shown, indacaterol maleate has obvious cyc...
Embodiment 3
[0037] Example 3 In vivo experiment: the inhibitory effect of indacaterol maleate on the growth of xenografted human colorectal cancer cells in nude mice.
[0038] (1) Select 18 6-week-old female nude mice (balb / c-nu / nu) (purchased from Southern Medical University), and randomly divide them into control group, experimental group 1 and experimental group 2, with 6 mice in each group, used to construct tumor models.
[0039] (2) HCT116 cells were digested, counted and resuspended in PBS buffer (pH=7.4, 0.01M), and the cell concentration in the cell suspension was 1×10 8 cells / mL, mix the cell suspension with Matrigel at a volume ratio of 1:1. Before the experiment, the nude mice were anesthetized, and the degree of anesthesia was evaluated through painless and painful stimulation to ensure that the nude mice were under anesthesia. HCT116 cells resuspended in PBS buffer and mixed with Matrigel were subcutaneously injected into nude mice with a micro-syringe with a 25G needle, a...
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