Application of curcuma zedoary extract in preparation of medicine for treating breast cancer bone metastasis
A technology of Curcuma curcuma extract and breast cancer, applied in the field of traditional Chinese medicine, can solve problems such as reports on the function of meso-Hannokinol, and achieve the effects of inhibiting bone metastasis and reducing bone damage
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Embodiment 1
[0020] Example 1 meso-Hannokinol inhibited the metastasis of breast cancer cells with high bone metastasis MDA-MB-231BO in the scratch test
[0021] Take the MDA-MB-231BO cells in the logarithmic growth phase, digest and centrifuge with trypsin, resuspend the cells in complete medium, and adjust the cell number to 3×10 with a cell counter. 5 cells / ml, inoculated into 6-well plates, 2ml per well, and cultured in an incubator. Use a 200 microliter pipette tip to draw a vertical scratch on the cell surface, then wash the cells twice with PBS to remove the scratched residual cells, add different concentrations of meso-Hannokinol serum-free medium for culture, Continue culturing for 24 hours until the cells adhere to the wall, observe the changes of cell migration after drug treatment at 0h and 24h under a microscope and take pictures, and calculate the scratch healing rate=(0h scratch width-24h scratch width) / 0h scratch width×100 %, which reflects the cell migration ability.
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Embodiment 2
[0023] Example 2 meso-Hannokinol inhibited the metastasis of breast cancer cells with high bone metastasis MDA-MB-231BO in Transwell experiment
[0024] MDA-MB-231BO cells were spread into the upper chamber of the transwell plate at 30,000 / well, and cultured in serum-free medium containing different concentrations of meso-Hannokinol, and culture medium containing 10% serum was added to the lower chamber of the transwell plate The base is used to induce the migration of tumor cells. After culturing for 24 hours, take out the small chamber and wash it twice with PBS, put the small chamber in paraformaldehyde fixative solution for 30 minutes, wash both sides with PBS again, and put the small chamber in crystal violet Stain the cells in the staining solution for 30 minutes, wash with PBS again, and gently wipe off the non-migrated cells on the upper layer of the chamber membrane with a cotton swab, photograph the migrating cells on the lower layer of the chamber membrane under a mi...
Embodiment 3
[0026] Example 3 Meso-Hannokinol inhibits the osteoclast differentiation of macrophage RAW264.7 cells induced by the conditioned medium of breast cancer cells with high bone metastasis MDA-MB-231BO
[0027] The culture medium of breast cancer cells with high bone metastasis MDA-MB-231BO was collected, filtered through a 0.22 μm aqueous phase filter, and mixed with medium containing 10% serum at a ratio of 1:1 to obtain breast cancer cells with high bone metastasis MDA-MB- Conditioned medium (CM) of 231BO. Spread RAW264.7 into a 12-well plate at 100,000 / well, and culture RAW264.7 cells for 48 hours with ordinary medium, conditioned medium, and conditioned medium containing meso-Hannokinol (50 μM) for 48 hours. Tartrate-resistant acid phosphatase detection kit detects the level of tartrate-resistant acid phosphatase (TRAP) in RAW264.7 cells. Tartrate-resistant acid phosphatase, a glycosylated metalloprotease, is considered to be an important indicator of osteoclastic activity i...
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