Application of eudesmane sesquiterpene lactone epimers in preparation of anti-breast cancer medicine
A technology of sesquiterpene lactones and epimers, which is applied in the field of chemistry and can solve unsolved problems such as the determination of the absolute configuration of epimers
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example 1
[0028] Example 1 Eucalyptane-type sesquiterpene lactone epimer anti-breast cancer cell MCF-7 activity test
[0029] 1. Experimental method: the breast tumor cell line MCF-7 was incubated with RPMI-1640 medium containing 10% calf serum at 37°C and 5% CO 2 cultured in an incubator. The MTT method was used to carry out the cell proliferation inhibition test. The main operation was to take the tumor cell lines in the logarithmic growth phase, digest them with 0.25% trypsin, and prepare 5×10 RPMI-1640 culture medium with 10% newborn calf serum. 4 pcs / ml -1 The cell suspension was inoculated in a 96-well plate with 180 μL per well. At 37°C, 5% CO 2 Cultivate for 8-10 hours under saturated humidity conditions, and when they adhere to the wall, add sample solution prepared with PBS to each well so that the final concentrations of the samples are 1, 10 and 100 μg / mL, respectively. For each concentration, 3 wells were paralleled, and after 44 hours of continuous culture, 50 μL MTT (...
Embodiment 2
[0035] Example 2 Observation of Sesquiterpene Lactone Epimer-Induced Morphological Changes in Breast Cancer Cell MCF-7 Cells under an Inverted Microscope
[0036] 1. Experimental method: Take the breast cancer cell MCF-7 in the logarithmic growth phase and press 1×10 6 cells / mL were inoculated into 6-well culture plates. The sample to be tested was dissolved in 0.1% DMSO to prepare a storage solution. Compounds 1 and 2 at the same concentration (10 μg / ml) were added to each administration group, and an equal volume of culture solution containing 0.1% DMSO was added to the vehicle control group. After culturing for 24 h, the morphological changes of the cells were observed and photographed under an inverted microscope. Such as figure 1 As shown, the effects of compounds 1 and 2 on the morphology of breast cancer cell MCF-7 were observed under an inverted microscope.
[0037] 2. Explanation of the experimental results: After 24 hours of drug treatment, the compound 2 group c...
Embodiment 3
[0038] Example 3 Detection of the level of reactive oxygen species ROS in breast cancer MCF7 cells
[0039] Add 3 × 10 per well in a six-well plate 5 MCF-7 cells with logarithmic growth were cultured for 24 hours, and culture medium with a concentration of 10 μg / mL of the sample to be tested was added and cultured for 36 hours, and then the fluorescent probe DCFH-DA was used to detect reactive oxygen species ROS. Add 1 mL of diluted DCFH-DA to each well, incubate at 37°C for 20 min in a cell culture incubator, wash the basal medium three times, and measure ROS levels by fluorescence intensity under an inverted fluorescence microscope. Or digest the cells, suspend in DCFH-DA solution with a final concentration of 10 μmol / L, incubate at 37° C. for 20 minutes in a cell incubator, and wash three times. The ROS content was detected by flow cytometry, the excitation wavelength was 502nm, the emission wavelength was 530nm, and the data was analyzed by FlowJo software. Experiments w...
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