Application of cnidium lactone and trastuzumab combined for preparing medicine for treating gastric cancer
A kind of osthole, combined preparation technology, applied in the field of medicine
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Embodiment 1
[0026] Example 1 The combination of osthole and trastuzumab has an in vitro inhibitory effect on gastric cancer N87 cells
[0027] Take N87 cells in good growth state, and use 1.5×10 cells per well 3 Spread evenly in 96-well cell culture plate, in the well cell culture plate, 37 ℃, 8% CO 2 Cultivate in the incubator for 24 hours; set trastuzumab monotherapy group, osthole monotherapy group and osthole combined trastuzumab group. Add trastuzumab (Tra, Trastuzumab) serially diluted from 50 μg / mL to each well, and repeat three times; add osthole (Ost, Osthole) serially diluted from 100 μM to each well; wait for drug treatment for 48 h After that, add CCK-8 reagent to develop color. The cell viability was calculated by reading the value at a wavelength of 450 nm on an iMark microplate reader.
[0028] Cell viability (%)=[(absorbance value of test group-absorbance value of blank group) / (absorbance value of control group-absorbance value of blank group)]×100
[0029] Experimenta...
Embodiment 2
[0033] Example 2 Combination of osthole and trastuzumab promotes apoptosis of N87 cells
[0034] Take N87 cells in good growth state to 5×10 6 / / well spread evenly in a 6-well plate. The cells in each well were subjected to the following four treatments: ① IgG (Control) (10 μg / mL); ② 10 μg / mL trastuzumab; ③ 40 μM osthole; ④ 10 μg / mL trastuzumab + 40 μM Osthole; the treatment condition is 37°C 8% CO 2 Incubate for 30 hours. Then stained with Annexin V and propidium iodide (PI), and the proportion of apoptotic cells was determined by flow cytometry.
[0035] Experimental results such as Figure 4 with Figure 5 as shown, Figure 4 is the N87 cell apoptosis level measurement chart, Figure 5 It is the proportion analysis chart of N87 cell apoptosis. The proportion of apoptotic cells in the combination group of trastuzumab and osthole was significantly higher than that in the group of trastuzumab alone or osthole alone. Therefore, the increase in the proportion of cell ap...
Embodiment 3
[0036] Example 3 Osthole and trastuzumab combined for inhibiting tumor cell growth in vivo
[0037] Female NOD-SCID mice were subcutaneously inoculated with gastric cancer N87 cells, and the tumor tissue reached 80 mm after inoculation of tumor cells. 3 -120mm 3 Size, intravenous injection of the following 4 groups of mice, 6 mice in each group: ① Human IgG (Control) (15 mg / kg); ② Trastuzumab (Trastuzumab) (15 mg / kg); ③ Osthole group (Osthole) (100 mg / kg); ④ Trastuzumab (15 mg / kg) + osthole (100 mg / kg) group (Trastuzumab + Osthole). The administration method of osthole was intraperitoneal administration every day, and the administration method of trastuzumab was once every three days for a total of 2 weeks. During the administration process, the tumor growth was observed every 2 days, and the tumor size and weight were measured to evaluate the antitumor effect of the drug. Experimental results such as Image 6 with Figure 7 shown. Image 6 The middle ordinate is the tum...
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