Application of N-hydroxyphthalimide compounds in preparation of anti-tumor medicine
A technology of hydroxyphthalimide and anti-tumor drugs, which is applied in the field of medicine, can solve the problems of unreported anti-tumor activity, poor pharmacokinetic properties, and large toxic and side effects, and achieve the purpose of inhibiting tumor growth and toxicity. Less side effects and convenient medication
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Embodiment 1
[0037] NHPI significantly inhibited mTORC1 and mTORC2 signaling pathways in different tumor cells.
[0038] 1. Cell lines: Human breast cancer BT-20 cells and human colon cancer Lovo cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Cells were placed at 37°C, 5% CO 2 Routine culture in an incubator with 10% calf serum, MEM and F12-K with a pH value of 7.4.
[0039] 2. Experimental method: take BT-20 and Lovo cells in the logarithmic growth phase, and make 3×10 5 / ml cell suspension was seeded in 6-well plate. After culturing for 24 hours, the original culture solution was aspirated after the cells adhered to the wall. Different concentrations of NHPI (purchased from Shanghai Shaoyuan Technology Co., Ltd.) were added to each well of the experimental group, and the same amount of culture solution without NHPI was added to the control group. After 24 hours, the cells were collected, the total cell protein was extracted, and subjected to SDS-P...
Embodiment 2
[0042] NHPI significantly inhibited the proliferation of different tumor cells.
[0043] 1. Cell lines: human breast cancer BT-20 cells, human colon cancer Lovo and HT-29 cells were purchased from Shanghai Cell Bank of Chinese Academy of Sciences.
[0044] 2. Experimental method: Take BT-20 cells, Lovo cells and HT-29 cells in the logarithmic growth phase, and make 1×10 4 / ml cell suspension was inoculated in a 96-well plate. After culturing for 24 hours, the original culture solution was aspirated after the cells adhered to the wall. Different concentrations of NHPI (purchased from Shanghai Shaoyuan Technology Co., Ltd.) were added to each well of the experimental group, and the same amount of culture solution without NHPI was added to each well of the control group. Set up 3 duplicate holes. After each group of cells was cultured for 48 hours, 20 μl of MTS (Promega, Madison, USA) was added to each well, and after another 1-2 hours of culture, the absorbance value at 490 nm...
Embodiment 3
[0047] NHPI blocks Lovo cell cycle at G 2 / M period.
[0048] 1. Experimental method: Take Lovo cells in the logarithmic growth phase and make 1×10 5 / ml cell suspension was seeded in 6-well plate. After culturing for 24 hours, the original culture solution was aspirated after the cells adhered to the wall. Different concentrations of NHPI (purchased from Shanghai Shaoyuan Technology Co., Ltd.) were added to each well of the experimental group, and the same amount of culture solution without NHPI was added to the control group. At the end of the treatment, the cells were collected by trypsinization and centrifuged, washed once with pre-cooled PBS, and fixed by adding 75% ethanol (pre-cooled at -20°C) overnight. Cells were collected by centrifugation and incubated with RNase at 37°C for 30 minutes, then stained with propidium iodide (PI, Sigma), and kept in the dark at room temperature for 30 minutes, and the cell cycle was detected by flow cytometry.
[0049] 2. Experimenta...
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