Use of Sichuang-Tibet amethystoidin A in preventing and treating acute lymphocyte leukaemia and cervical cancer
A technology of Sichuan-Tibetan incense tea A, acute lymphocytes, which can be used in the fields of biotechnology and medicine and can solve problems such as drug resistance
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Embodiment 1
[0101] Example 1 cell treatment
[0102] The human T-cell leukemia cell line Jurkat and the human B-lymphoma cell line Raji were treated with 10% heat-inactivated fetal bovine serum (FBS, Gibco BRL, Gaithersburg, ML), penicillin (100 IU / ml) and streptomycin (100 μg / ml) in RPMI 1640 medium (Sigma-Aldrich, St Louis, MO), placed at 37°C, containing 5% CO 2 and 95% air incubator. Hela (cervical carcinoma) cells were obtained from the American Standard Biological Collection (ATCC, Manassas, VA) and cultured in Dulbesso containing 10% heat-inactivated fetal calf serum on Petri dishes or on Lab-Tek II slides (Fisher Scientific). 's modified Eagle's medium (DMEM), placed at 37 ° C, containing 5% CO 2 and 95% air incubator. Paclitaxel-resistant Jurkat cells were supplemented with 20 nM paclitaxel (Sigma-Aldrich, St Louis, MO) every other day and maintained for a long time. In the experiment, the cells were 2 × 10 5 Cells / ml were seeded and treated with corresponding concentra...
Embodiment 2
[0103] Example 2 Flow Cytometry Analysis
[0104] For flow cytometry analysis of DNA content, collect 10 6 Cells were rinsed and fixed overnight at -20°C with 75% cold ethanol. Cells were treated with 100 μg / ml RNase A dissolved in Tris-HCl buffer (pH 7.4) and stained with 25 μg / ml propidium iodide (PI, Sigma-Aldrich, St Louis, MO), and samples were analyzed by flow cytometry (FACSCalibur, BD Biosciences, San Jose, CA, USA) CellQuest Pro software (BD Biosciences) was used. 10,000 cells were selected for DNA content analysis.
Embodiment 3
[0105] Example 3 co-immunoprecipitation
[0106] Equal amounts of cell lysates from different treatments were incubated overnight at 4°C with the rabbit polyclonal antibody Cdc20. Protein A Sepharose beads (Santa Cruz, CA) were added for 2 hours and incubated at 4°C. Rabbit IgG (Santa Cruz, CA) was used as a negative control before normal immunization. Immunoprecipitates were collected, rinsed with lysis buffer, and dissolved in 2× sodium dodecyl sulfate (SDS) for western blot analysis.
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