Application of losmapimod to non-small cell lung cancer
A technology for non-small cell lung cancer and drug-resistant cells, which can be used in medical preparations containing active ingredients, respiratory diseases, organic active ingredients, etc., and can solve the problems of lack of further application and lack of research reports.
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Embodiment 1
[0047] This example mainly introduces the relationship between the p38 phosphorylation activation pathway and the production of tetraploid cells, as well as the relationship between Losmapimod and tetraploid cells. The related experiments are briefly introduced as follows.
[0048] Before introducing the specific experiments, some detection methods and experimental methods in the following experiments are briefly introduced as follows.
[0049] (1) Cell Viability Detection Method
[0050] When the cells are subcultured, digest the adherent cells with trypsin, resuspend the cells with the corresponding medium after centrifugation, adjust the cell concentration, and inoculate 3000 cells per well into a 96-well plate with a volume of 100 μL per well;
[0051] After 24 hours, add corresponding concentration of drug treatment (the drug is finally dissolved in 100 μL medium);
[0052] After 72 hours, add 20 μL of 0.3 mg / mL thiazolium blue (MTT, 3-(4,5-dimethylthiazole-2)-2,5-diphen...
Embodiment 2
[0086] Through the MTT cell activity detection experiment (refer to Example 1 for the experimental method) and the soft agar experiment, this example mainly tests the changes in the proliferation ability and clone formation ability of tumor cells after Losmapimod is used in combination with gefitinib. Before introducing the specific experiment, the soft agar experimental method is briefly introduced as follows.
[0087] experiment
[0088] Prepare 1.2% agarose, after autoclaving, place it in a 44°C water bath to keep the agarose in a melted state;
[0089] Prepare 2×1640, filter it with a 0.22 μm filter membrane into a sterilized container, and store it in a refrigerator at 4°C;
[0090] Prepare the lower layer glue, taking 50ml as an example: 20ml of 2×1640, 7.5ml of fetal bovine serum, 2.5ml of 1×PBS, 20ml of 1.25% agar, 5μL of gentamicin (50mg / ml), after preparation , placed in a 44°C water bath to keep warm;
[0091] Prepare 1640 medium with 15% FBS, taking 50 ml as an ...
Embodiment 3
[0105] On the basis of the above Examples 1 and 2, the inventors further carried out in vivo experiments on xenografted lung cancer mice resistant to gefitinib derived from patient tumor tissue (the tumor in this case was provided by Henan Cancer Hospital). The detailed experimental process The introduction is as follows.
[0106] The gefitinib-resistant non-small cell lung cancer tissue was inoculated subcutaneously into 40 mice, and the mice were divided into 4 groups (10 mice in each group); when the tumor volume in the mice was about 25mm 3 At that time, the four groups of mice were treated as follows:
[0107] 1. Control group (5% DMSO, 50% normal saline, 50% PEG400);
[0108] 2. 50mg / kg gefitinib treatment group;
[0109] 3. Losmapimod treatment group at 12 mg / kg;
[0110] 4. Co-treatment group of 50 mg / kg Gefitinib and 12 mg / kg Losmapimod (gastric administration);
[0111] Body weight and tumor volume (length × width) of the mice were measured twice a week 2 ×0.5)...
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