Antineoplastic combined drug, and applications thereof in preparation of anticancer drugs
An anti-cancer drug and anti-tumor technology, applied in the field of biomedicine, can solve the problems of poor selectivity, drug resistance, toxic and side effects, etc.
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Embodiment 1
[0036] Example 1 Effects of Medication Alone and Combined Medication on Different Normal Cell Lines
[0037] 1.1 Materials and Instruments
[0038] Test drugs: chlorogenic acid, erlotinib, gefitinib, icotinib, afatinib, trastuzumab, lapatinib, nimotuzumab, cetuximab .
[0039] Tested cell lines: human lung cancer cell line A549, human breast cancer cell line MDA-MB-453, human nasopharyngeal carcinoma cell line CNE-1, human normal lung epithelial cell BEAS-2B, human normal breast cell MCF10A, human normal nasal cavity Epithelial cells RPMI2650.
[0040] 1.2 Experimental method
[0041] Take a bottle of cells in the logarithmic growth phase, wash them twice with PBS, digest with 0.25% trypsin, and prepare a single cell suspension with RPMI-1640 culture medium containing 10% calf serum for counting. Inoculate in a 96-well culture plate under sterile conditions, and the number of inoculated cells is 5×l0 3 / hole. cells at 37°C, 5% CO 2 After incubation for 24 h in the incub...
Embodiment 2
[0066] Example 2 Chlorogenic acid reverses the drug resistance of human lung adenocarcinoma cell PC9 to gefitinib in vitro
[0067] 2.1 Materials and Instruments
[0068] Test drug: chlorogenic acid; gefitinib
[0069] Tested cell lines: PC9 cells, PC9 / ZD is a self-screening gefitinib-resistant cell line
[0070] 2.2 Experimental method
[0071] 2.2.1 Cultivation of drug-resistant cell lines
[0072] Human lung adenocarcinoma cell line PC9 cells were exposed to gefitinib (200nmol / L) for up to 3 months, and the gefitinib-resistant cell clones were selected to become PC9 / ZD cell lines. These two kinds of cells respectively have the ability of stable drug resistance to gefitinib. PC9 cells and PC9 / ZD cells were grown on the wall at 25cm 2 Culture flasks were cultured in DMEM medium containing 10% fetal bovine serum in 5% CO 2 , and incubated in a 37°C incubator. Observe the cells every day. When the cells grow to cover 70%-80% of the bottom of the bottle, they are subcultu...
Embodiment 3
[0089] Example 3 Synergistic and attenuating effects of chlorogenic acid and EGFR-targeted drug combination on tumor inhibition in vivo
[0090] 3.1 Materials and Instruments
[0091] Test drug: chlorogenic acid; afatinib, erlotinib, panitumumab, nimotuzumab
[0092] Tested cell lines: G422 cells, Lewis mouse lung cancer cell lines
[0093] Test animals: BABLc mice, C57BL / 6 mice
[0094] 3.2 Experimental method
[0095] 3.2.1 Establishment of experimental animal tumor models
[0096] Collect the cells in the logarithmic growth phase, centrifuge at 1000rpm for 5min, wash the cell sediment twice with PBS, and adjust the cell concentration to 1x10 with serum-free medium after counting 7 / m1, sent to the animal room under sterile conditions. Under sterile experimental conditions, mice were subcutaneously injected with 1x10 7 / m1 cells, 0.1ml per mouse, obvious skin mounds appeared in the injection area, and large rice grain nodules appeared in the left armpit of the nude mic...
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