Application of Jinfei capsule in preparation of anti-breast cancer drugs
A technology for breast cancer, breast cancer cells, applied in the field of biopharmaceuticals
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Embodiment 1
[0036] Cell Viability Test
[0037] Preparation of sample solution: The contents of Jingan capsules were prepared with physiological saline to prepare sample solutions with concentrations of 0.125mg / ml, 0.25mg / ml, 0.5mg / ml, 1mg / ml, and 2mg / ml.
[0038] Preparation of MTT solution: Weigh 0.2g of MTT powder, add 40ml of sterilized double distilled water, dissolve at 60°C, the concentration is 5mg / ml, sterilize by 0.2μm filter membrane, and store at 4°C.
[0039] Cell lines: human breast cancer cells MDA-MB-231, MDA-MB-468 cells and mouse breast cancer cell 4T1, all of which are reserved by the Key Laboratory of Natural Product Chemistry, Chinese Academy of Sciences, Guizhou Province. cells at 37°C, 5% CO 2 Cultured in DMEM medium containing 5% FBS in an incubator.
[0040] Collect logarithmic phase cells, MDA-MB-231 cells, MDA-MB-468 cells and 4T1 cells according to 6×10 3 Cells / well were inoculated in 96-well plates and cultured overnight. After the cells adhered to the wall, ...
Embodiment 2
[0046] Cell Growth Curve Determination
[0047] Collect logarithmic phase MDA-MB-231 cells and MDA-MB-468 cells respectively, according to 6×10 3 The density of cells / well was inoculated in 96-well plate, cultivated overnight, and after the cells adhered to the wall, different concentrations of sample solutions (0.25mg / ml, 0.5mg / ml, 1mg / ml, 2mg / ml) were added for treatment, respectively. OD 490nm was measured at 0h, 24h, 48h, 72h, and 96h, and the growth curve was drawn.
[0048] The result is as image 3 as well as Figure 4 As shown, compared with the control group, different concentrations of sample solutions can inhibit the proliferation of MDA-MB-231 cells and MDA-MB-468 cells, and with the increase of concentration, MDA-MB-231 cells and MDA-MB The growth rate of -468 cells gradually slowed down, indicating that the sample solution could inhibit the proliferation of MDA-MB-231 and MDA-MB-468 cells in a concentration- and time-dependent manner.
Embodiment 3
[0050] Apoptosis detection
[0051] MDA-MB-231 cells were divided into 4 × 10 5 The amount of cells / well was inoculated in a 60mm round petri dish for culture. After the cells adhered to the wall, different concentrations of sample solutions (0.5mg / ml, 1mg / ml, 2mg / ml) were added to act for 24h or 48h, and then collected cell. Wash 3 times with PBS, add 500 μl 1×Binding Buffer to resuspend the cells, add 5 μl AnnexinV-FITC and 10 μl propidium iodide, incubate at room temperature in the dark for 15 min, and calculate the cell apoptosis rate by flow cytometry.
[0052] The apoptosis experiment of MDA-MB-468 cells was the same as that of MDA-MB-231 cells.
[0053] The result is as Figure 5 As shown, Jingan Capsule has a significant effect on the apoptosis of human breast cancer cell MDA-MB-231.
[0054] The result is as Figure 6 As shown, Jingan Capsule has a significant effect on the apoptosis of human breast cancer cell MDA-MB-468.
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