Application of Fenghuang Dancong tea extract in preparation of anti-breast-cancer drugs
A technology of anti-breast cancer and Dancong tea, which is applied in the field of medicine and food, can solve problems such as no public reports, and achieve great safety, good killing effect, and the effect of improving the body's immunity
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Embodiment 1
[0039] Example 1 Cell Viability Experiment
[0040] A MTT method was used to measure the effects of the water extract of Fenghuang Dancong tea on four human malignant tumor cells (human breast cancer cell MDA-MB231, human colon cancer cell SW480, human cervical cancer cell Hela, human liver cancer cell HepG2) and one The method for the toxicity of normal cells (human hepatocyte L02) (tumor cells and normal cells are purchased from the U.S. Model Culture Collection Bank), mainly includes the following steps:
[0041] (1) Dissolve the tea water extract of Fenghuang Dancong tea in PBS buffer solution, put it on an ultrasonic instrument and sonicate for 1-2 minutes, and prepare a 5 mg / mL stock solution for later use;
[0042] (2) Different tumor cells and normal cells in the logarithmic growth phase were taken at a density of 2×10 4 cells / mL were inoculated in 96-well plates (100 μL / well), and allowed to grow on the wall for 24 hours. Then add 100 μL of drugs with different conc...
Embodiment 2
[0044] Example 2 Flow cytometry detection of the influence of Fenghuang Dancong tea water extract on the distribution of tumor cell cycle
[0045] A method for detecting the influence of Phoenix Dancong tea water extract on the cell cycle distribution of human breast cancer MDA-MB231 by flow cytometry, mainly comprising the following steps:
[0046] (1) Dissolve the tea water extract of Fenghuang Dancong tea in PBS buffer solution, put it on an ultrasonic instrument for 1-2 minutes, and prepare a 5 mg / mL solution for later use;
[0047] (2) MDA-MB231 cell density is 2×10 4 / mL in a 6mL culture dish for 24 hours, after the cells are completely adhered to the wall, add the water extract of Fenghuang Dancong tea leaves with different concentrations prepared by using the stock solution prepared in step (1), incubate for 72 hours, and collect the cells. Fix with pre-cooled 70% ethanol overnight at -20°C. Centrifuge at 1500rpm for 10min, discard the culture medium, turn it upside ...
Embodiment 3D
[0049] Embodiment 3DHE probe detects intracellular reactive oxygen species (ROS) level
[0050] A method using DHE probes to detect the influence of Phoenix Dancong tea water extract on the level of reactive oxygen species (ROS) in human breast cancer MDA-MB231 cells, mainly comprising the following steps:
[0051] (1) Dissolve the tea water extract of Fenghuang Dancong tea in PBS buffer solution, put it on an ultrasonic instrument and sonicate for 1-2 minutes, and prepare a 60 μg / mL solution for later use.
[0052] (2) MDA-MB231 cell density is 2×10 4 / mL in a 96-well plate for 24 hours, add Phoenix Dancong tea water extracts with concentrations of 60 μg / mL, 30 μg / mL, and 15 μg / mL, and incubate for 2 hours, remove the medium in each well, and add 100 μL of 10 μM DHE Probes (reagents purchased from Sigma) were incubated for 30 min. The excitation and emission wavelengths of the fluorescent microplate reader were set to 300nm and 610nm respectively, and the fluorescence absor...
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