Novel application of artesunate
A new technology of artesunate, applied in the field of lung cancer treatment, can solve the problems of drug resistance and limited application of molecular targeted drugs, and achieve the goal of reversing erlotinib resistance, solving drug resistance and improving the prognosis of lung cancer Effect
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experiment example 1
[0026] Antitumor activity of artesunate on human lung adenocarcinoma PC9 cells in vitro
[0027] 1 Materials and methods
[0028] 1.1 Main experimental materials
[0029] 1.1.1 Cell lines
[0030] Human lung adenocarcinoma PC9 cells were kindly donated by Professor Chen Zhengtang, Army Tumor Institute, Xinqiao Hospital, Third Military Medical University.
[0031] 1.1.2 Main reagents
[0032]
[0033] 1.1.3 Main materials and equipment
[0034]
[0035]
[0036] 1.2 Experimental method
[0037] 1.2.1 Recovery, culture, subculture and cryopreservation of cells
[0038] Tumor cell recovery and culture
[0039] Take out the cryopreservation tube containing the human lung adenocarcinoma PC9 cell line from the liquid nitrogen tank, put it into a beaker filled with warm water at 37°C-40°C and shake it quickly, so that the frozen cell solution can be quickly dissolved within 30s, and the cells Transfer to a sterile centrifuge tube, add 5ml of RPMI-1640 culture medium (...
experiment example 2
[0107] In Vitro Experiment of Artesunate Reversing Erlotinib Resistance in PC9 / ER Cells
[0108] 1 Materials and methods
[0109] 1.1 Main experimental materials
[0110] 1.1.1 Main reagents
[0111] LY294002 was first formulated into a 20nmol / L mother solution (dissolved in DMSO), stored in a -20°C refrigerator for a long time, and the working solution was prepared with fresh cell culture medium before use as required by the experiment.
[0112] 1.2 Experimental method
[0113] 1.2.1 The experimental methods of cell recovery, culture, subculture and cryopreservation are the same as before.
[0114] 1.2.2 The cck8 kit was used to detect the effect of ART on the proliferation ability of PC9 / ER cells.
[0115] PC9 cells and PC9 / ER cells in the logarithmic growth phase were mixed at 5×10 per well 3 Cell density, seeded in 96-well plate, 100 μl per well, placed at 37°C, 5% CO 2 After culturing in the incubator for 24 hours, ART was added to make the final concentrations of 6...
experiment example 3
[0138] In Vivo Experiment of Artesunate Reversal of Erlotinib Resistance in PC9 / ER Cells
[0139] 1 Experimental method
[0140] 1.1 Establishment of experimental animal tumor models
[0141] Select a flask of PC9 / ER cells in the logarithmic growth phase, then digest the cells with 0.25% trypsin 1-2ml containing 0.025% EDTA, observe under an inverted microscope, and quickly add a small amount of 10% FBS when the cells withdraw their protrusions and become round Gently blow and blow the digested cells repeatedly to separate them from the culture flask wall and disperse them into a cell suspension. Then transfer the cells into a new centrifuge tube and centrifuge on a centrifuge (1000rpm, 5min). The white cell pellet was washed 2-3 times with PBS solution, counted with a cell counting plate, and then the cell concentration was adjusted to 1×10 with fresh culture medium (without FBS). 7 / ml, sent immediately to the SPF grade animal breeding room under sterile conditions. Injec...
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