Anti-tumor combined drug and use thereof in preparing anti-cancer drug
An anticancer drug and antitumor technology, applied in the field of biomedicine
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Embodiment 1
[0034] Example 1 MTT determination of proliferation inhibition of combination medication
[0035] 1.1 Materials and instruments
[0036] Tested drugs: chlorogenic acid, sorafenib, sunitinib, axitinib, pazopanib, bevacizumab.
[0037] Test cell lines: human lung cancer cell line A549, human kidney cancer cell line 786-0, human normal lung epithelial cell BEAS-2B, human embryonic kidney normal cell CCC-HEK-1
[0038] 1.2 Experimental method
[0039] Take a bottle of logarithmic growth phase tumor cells, wash twice with PBS, digest with 0.25% trypsin, and make a single cell suspension with RPMI-1640 culture medium containing 10% calf serum for counting. Inoculate 100μl of cells on 96-well culture plate under aseptic conditions, and the number of inoculated cells is 5xl0 3 / hole. Cells at 37℃, 5% CO 2 After 24h incubation in the incubator, the medium was changed.
[0040] The cells were divided into negative control group, single medication group, chlorogenic acid group and combination med...
Embodiment 2
[0061] Example 2 Anti-tumor animal test of combination medication
[0062] 2.1 Materials and methods
[0063] Test drugs: chlorogenic acid; VEGFR tyrosine kinase inhibitors: sorafenib, sunitinib, axitinib, pazopanib, bevacizumab
[0064] Test cell lines: lung cancer cell line A549 cells, human kidney cancer cell line 786-0
[0065] Tested animal: BABL / C-nu mouse, ♀, weighing 18-22g;
[0066] 2.2 Test method
[0067] 2.2.1 Establishment of experimental animal tumor model
[0068] Collect the logarithmic growth phase cells, centrifuge at 1000 rpm for 5 min, wash the cell sediment with PBS twice, after counting, adjust the cell concentration to 1x10 with serum-free medium 7 / m1, sent to the animal room under aseptic conditions. Under sterile experimental conditions, 1x10 was injected subcutaneously into the right armpit of the mouse 7 / m1 cells, 0.1ml each, obvious skin hills appeared in the injection area, and a large rice nodule appeared on the left axillary of the mouse one week later, i...
Embodiment 3
[0104] Example 3 Animal test of chlorogenic acid in the preparation of anti-multidrug resistance combination drugs
[0105] 3.1 Materials and methods
[0106] Test drug: chlorogenic acid; VEGF tyrosine kinase inhibitor: sorafenib, pazopanib.
[0107] Tested cell line: lung cancer drug-resistant cell line A549. The A549 cell line was induced by the sorafenib concentration gradient method, and established after clonal screening, and cultured before the experiment.
[0108] The human renal cancer drug-resistant cell line 786-0 was induced by the pazotinib concentration gradient method to induce the A549 cell line, which was established through clonal screening and cultured before the experiment.
[0109] Tested animal: BABL / C-nu mouse, ♀, weighing 18-22g;
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