The application of etoside as antitumor drug
A technology of etoside and drugs, which is applied in the application field of etoside as an anti-tumor drug, and can solve the problems of large side effects, easy drug resistance, easy recurrence, etc.
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Embodiment 1
[0048] The embodiment of the present invention provides the effect of etoside as a broad-spectrum anti-tumor drug on the proliferation activity of various tumor cells in vitro, specifically as follows:
[0049] 1. Experimental method
[0050] 1.1 Experimental cell lines
[0051] EA.hy926 (human umbilical vein cell fusion cell line), from the State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine;
[0052] MYO (primary cardiomyocyte cell line of neonatal rats) was obtained from the State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine;
[0053] U2OS (human osteosarcoma cell line), from the State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine;
[0054] A2780 (human cervical cancer cell line), from the State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine;
[0055] Hela (human ovarian cancer cell...
Embodiment 2
[0074] The embodiment of the present invention provides the effect of etoside as an anti-osteosarcoma drug on the cell cycle of osteosarcoma.
[0075] 1. High-content cell imaging system to detect the effect of etoside on osteosarcoma cell cycle
[0076] The cells in the above 1.2 were inoculated in 96-well cell culture plates with 1% FBS, 8000 cells per well, and after 24 hours of adherent culture, the control group was given DMEM, and the administration group was given samples containing etoside. 100nmol / L, 10nmol / L, 1nmol / L DMEM. Each group had 3 replicate wells, and the cells were stored at 37°C, 5% CO 2 , cultured under saturated humidity conditions, and after 24 hours of action, perform immunofluorescence staining: label EDU with fluorescent dye Alexa488 (green) to identify S-phase cells; use phosphorylated histone PHH3 and its fluorescent dye Alexa647 to label (red) Cells in the M phase; use the fluorescent dye Hochest33342 (blue) to mark all the nuclei to determine t...
Embodiment 3
[0084] The embodiment of the present invention provides the effect of etoside as an anti-osteosarcoma drug on the apoptosis of osteosarcoma cells.
[0085] 1. Flow cytometry detection of the effect of etoside on the apoptosis of U2OS cells
[0086] The 3rd to 9th generation U2OS cells were selected, and the DMEM high-glucose medium containing 10% FBS was used at 37°C, 5% CO 2 Carry out routine culture in the incubator, and when the fusion reaches about 80%, change to 1% FBS and synchronize for 24 hours. , 1nmol / L DMEM, incubate for 24 hours; suck off the supernatant of the medium, add an appropriate amount of PBS to rinse, shake the cell culture vessel gently, let the PBS cover the entire vessel surface, and then aspirate the PBS as clean as possible.
[0087] Add an appropriate amount of trypsin, incubate at 37°C, take it out every 1 minute or so, gently shake the cell culture vessel or pat the side wall of the cell culture vessel to observe whether the cells float smoothly....
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