Application of curcumin in preparation of thyroid carcinoma therapeutic agent
A technology for thyroid cancer and thyroid cancer cells, which is applied in the medical field of curcumin, can solve the problem that curcumin has no report on the effect of thyroid cancer cells, and achieve a good intervention effect and the effect of promoting apoptosis
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Embodiment 1
[0028] Example 1: Curcumin inhibits the proliferation of primary culture or thyroid cancer cell lines.
[0029] 1. Experimental materials
[0030]The experimental methods and reagents used in the following examples are conventional methods and reagents unless otherwise specified.
[0031] 1.1 Cell lines: The papillary carcinoma cell line K1, the follicular carcinoma cell line FTC-133, and the undifferentiated carcinoma cell line 8505C used in this experiment were all purchased from the European Cell Bank. FTC-133 cells were cultured with DMEM and Hams F-12 medium 1:1 (v / v); K1 cells were cultured with DMEM medium, Hams F-12 medium and MCDB105 medium 2:1:1 (v / v) For culture, 8505C cells were cultured using MEM medium. The above three cell lines were cultured in the corresponding medium supplemented with 10% fetal bovine serum (Hangzhou Sijiqing).
[0032] 1.2 Primary cell culture process: Take an appropriate amount of thyroid cancer tissue, paracancerous or contralateral nor...
Embodiment 2
[0038] Example 2: Growth inhibitory effect of curcumin on primary culture or thyroid cancer cell lines.
[0039] 1. Experimental materials
[0040] AnnexinV-FITC (fluorescein isothiocyanate-labeled phospholipid-binding protein 5) cell apoptosis detection kit, BD Pharmingen, Cat. No. 556547.
[0041] 2. Experimental method
[0042] Apoptosis detection was performed according to the instructions of the AnnexinV-FITC Apoptosis Detection Kit from BD Company. Cells were resuspended in 1× binding buffer and adjusted to 1×10 6 / mL, add 5 μL of Annexin V-FITC and 5 μL of PI solution to every 100 μL of cell suspension, and store at room temperature in the dark for 15 minutes. Add 400 μL of 1× Binding Buffer to each tube prior to flow cytometry analysis. Flow cytometry, FL1 and FL2 channels were used to detect AnnexinV and PI positive cells, and statistically analyze the cell apoptosis rate.
[0043] 3. Experimental results
[0044] Curcumin reacted with K1 or FTC-133 cells for 24...
Embodiment 3
[0045] Example 3: Curcumin promotes the cleavage of PARP in thyroid cancer cells and inhibits the expression of anti-apoptotic protein Bcl-2.
[0046] 1. Experimental materials
[0047] PARP primary antibody, Beyotime Biotechnology Company, 1:1000 dilution; Bcl-2 primary antibody, Beyotime Biotechnology Company, 1:1000 dilution; horseradish peroxidase-labeled goat anti-rabbit secondary antibody, Beyotime Biotechnology Company , 1:1000 dilution. ECL chemiluminescence kit, Biyuntian Biotechnology Company.
[0048] 2. Experimental method
[0049] The expression of apoptosis proteins in cells was detected by Western blot. K1 and FTC-133 cells in 4×10 5 Cells / mL were inoculated into 6-well cell culture plates, 1 mL per well, 37°C, 5% CO 2 After culturing overnight, add different concentrations of curcumin (10-50 μM) or solvent control group treatment, collect cells after culture, centrifuge at 6000 rpm for 5 minutes, wash twice with PBS, add 25 μL cell lysis buffer [150 mM NaC...
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