Application of Nifeviroc in preparing antitumor drug
An anti-tumor drug and tumor technology, which is applied in the field of medicine, can solve the problems of poor tumor treatment effect and many toxic reactions, achieve the reduction of directional migration ability and non-directional migration ability and in vitro invasion ability, significant effect, inhibition of invasion and transfer effect
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Embodiment 1C
[0030] Embodiment 1CCK-8 method detects cell proliferation
[0031] Cells in good growth state (non-small cell lung cancer A549 cells, non-small cell lung cancer NCI-H520 cells, human bronchial epithelial HBE cells, liver cancer HepG2 cells and breast cancer MCF-7 cells) were taken, digested with 0.25% trypsin, and mirrored. After observing the retraction of the cells, the digestion was terminated, centrifuged at 1000rpm for 5min, and the cells were collected. After staining with trypan blue, the cells were counted by a cell counter, and the cell density was adjusted to 4×10 4 cells / ml, seeded in 96-well plate, 100 μl per well (3×10 3 cells), set CO 2 cultured in a constant temperature incubator. The control group and the administration group were set up (the concentrations of Nifeviroc were 1.875, 3.75, 7.5, 15, and 30 μM respectively), and 5 multiple wells were set in each group. After the cells adhered to the wall, the original culture solution was discarded, and the admi...
Embodiment 2
[0034] Example 2 Annexin V-FITC / PI double staining method to detect cell apoptosis
[0035] Cells in good growth state (non-small cell lung cancer A549 cells, non-small cell lung cancer NCI-H520 cells, human bronchial epithelial HBE cells, liver cancer HepG2 cells and breast cancer MCF-7 cells) were digested with 0.25% trypsin to prepare Single cell suspension at 1 x 10 5 Inoculate in a 6-well culture plate at a density of 1 / well and store at 37°C, 5% CO 2 Incubator cultivation. After the cells adhered to the wall, the original culture medium was discarded, and the same volume of different concentrations of Nifeviroc (0, 1.875, 3.75, 7.5, 15, 30 μM) was added for intervention in groups, with 3 replicate wells for each group. After being treated with different concentrations of Nifeviroc for 48 hours, the cells were digested with EDTA-free trypsin and collected by centrifugation; the cells were washed three times with pre-cooled PBS buffer at 4°C, centrifuged at 1000 rpm for ...
Embodiment 3
[0039] Example 3 Cell clone formation experiment
[0040] Non-small cell lung cancer A549 cells in good growth state were digested with 0.25% trypsin to prepare a single cell suspension, seeded in a 24-well culture plate at a density of 100 cells / well, and placed at 37°C and 5% CO 2 Incubator cultivation. After the cells adhered to the wall, the original culture medium was discarded, and the same volume of different concentrations of Nifeviroc (0, 1.875, 3.75, 7.5, 15, 30 μM) was added for intervention in groups, with 3 replicate wells for each group. After 48 hours of Nifeviroc at different concentrations, change the fresh medium to continue culturing for 2-3 weeks, and change the medium every 72 hours; when clones visible to the naked eye appear in the culture plate, terminate the culture, discard the cell culture medium, and carefully soak in PBS. Wash twice, fix at room temperature for 15 minutes; remove the fixative, and stain with crystal violet for 30 minutes; slowly w...
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