Application of salinomycin in preparing medicament for resisting various human malignant tumors
A technology for malignant tumors and salinomycin, applied in the field of chemical medicine, can solve the problem of unclear influence of stem cells on inhibition
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Embodiment 1
[0010] Embodiment 1: In vitro anti-tumor cell proliferation experiment of salinomycin
[0011] The selected cell lines include human lung cancer cell A549, skin cancer cell A-431, pancreatic cancer cell BxBCO, colon cancer cell Caco2, colorectal cancer cell HT-29, liver cancer cell HepG2, tongue cancer cell SCC-9, renal cell carcinoma Cell 786-O, brain metastatic prostate cancer cell DU-145, etc. Take tumor cells in the logarithmic growth phase to make a certain concentration of cell suspension and inoculate them in a 96-well plate. After the cells are cultured for 48 hours, take out the culture plate, add 50ul of pre-cooled 50% trichloroacetic acid to each well, and let it stand After 5 minutes, move it into a 4°C refrigerator and let it stand for 1 hour, take it out and wash it 5 times with deionized water, and dry it in the air. After it is completely dry, add 100ul of 0.4% SRB prepared with 1% acetic acid to each well, and pour it out after 20 minutes of staining Dye solu...
Embodiment 2
[0021] Example 2: Effect of Salinomycin on Human Lung Cancer Cell Line A549 Stem Cells
[0022] 1) Isolation of lung cancer stem cells from human lung cancer cell line A549:
[0023] A549 cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). These cells were cultured in medium recommended by ATCC and tested in logarithmic growth phase. The ALDEFLUOR kit (StemCell Technologies, Durham, NC, USA) was used to react with human ALDH1A1 as much as possible to determine cancer stem cells. In simple terms, cells expressing fluorescent marker ALDH1A1 were detected and sorted by flow cytometry ( BD Biosciences, San Jose, CA, USA). Specifically, ALDH1A1 activation is based on the expression of the ALDEFLUOR inhibitor diethylaminobenzaldehyde (DEAB). Data were analyzed using Quest software (BD Biosciences).
[0024] [Flow cytometry detection and sorting of cells positive for the expression of the fluorescent marker ALDH1A1 are shown in Figure 2-1] ...
Embodiment 3
[0027] Example 3: The animal experiment on the effect of salinomycin on human lung cancer A549 stem cells is in the process of experimentation.
[0028] Nod-scid immune triple-deficient mice were purchased from Shanghai Slack Experimental Animal Co., Ltd. 5000 ALDH-positive A549 cells were suspended in 1 ml of Matrigel medium (Viglass Biotechnology (Beijing) Co., Ltd.), diluted 1:2 in DMEM, and injected subcutaneously into NOD-SCID mice. Tumor formation was detected by palpation. Salinomycin injection was given 24 hours later. The use of animals was carried out in accordance with the relevant national regulations. Salinomycin (5 mg / kg / day) or paclitaxel (5 mg / kg / day) was injected intraperitoneally once a day to 6 mice in each group for a total of four weeks.
[0029] In summary, salinomycin has the potential and prospect of being developed into an antitumor drug.
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