Application of platycodon saponin d in the preparation of antitumor drugs
A technology of tumor drug, platycodon saponin, applied in the medical field, can solve the problem of destroying the human immune system and achieve the effect of reducing adverse reactions
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Embodiment 1
[0011] Example 1: Implementation steps of the effect of platycodon saponin D on the viability of osteosarcoma cells and human osteoblast hFOB cells.
[0012] The osteosarcoma cell line Saos2-lung was cultured in vitro, which is a subpopulation of osteosarcoma cells with high metastatic potential isolated from lung metastases of tumor-bearing nude mice, and was labeled with luciferase. The parental cells were the most common Saos2 osteosarcoma cells. Simultaneously culture hFOB cells. The Saos2-lung and hFOB cells grown in logarithmic phase were digested with trypsin and centrifuged at 300g for 5min. After centrifugation, the supernatant was removed, and the cells were resuspended in complete medium and counted individually. The two kinds of cells were seeded into 96-well plates at the number of 5000 / well. The next day after the cells were plated, platycodon saponin D of corresponding concentration was added, and the concentrations of platycodon saponin D were 0, 5, 10 and 20...
Embodiment 2
[0015] Example 2: Implementation steps of the effect of Platycodon saponin D on apoptosis of osteosarcoma cells.
[0016] 1. Apoptosis Induction
[0017] Saos2-lung cells grown in logarithmic phase were taken, digested and centrifuged. After centrifugation, the supernatant was removed, and the cells were resuspended with DMEM complete medium and counted. cells in 2 x 10 5 The number of cells / well was planted in a 6-well plate, the cells were divided into three groups, the concentrations of Platycodon saponin D were 0, 10 and 20 μM, respectively, and three replicate holes were set in each group. After the cells adhered to the wall, platycodon saponin D was added to induce apoptosis, and apoptosis induction was carried out in serum-free medium.
[0018] 2. Apoptosis Detection
[0019] Cells were collected according to the cell subculture method, and EDTA-free trypsin should be used for trypsin. Collect the cell suspension into a flow tube. After centrifuging at 1500rpm for...
Embodiment 3
[0021] Example 3: Implementation steps of the effect of Platycodon saponin D on the growth and metastasis of osteosarcoma in vivo.
[0022] 1. Establishment of osteosarcoma model
[0023] Take the Saos2-lung osteosarcoma cells growing in the logarithmic phase, then digest and centrifuge the cells, resuspend the osteosarcoma cells with PBS solution and count them with a cell counter, observe the viability of the cells with trypan blue staining, and then centrifuge again to pellet the cells . Resuspend the cells in PBS solution to make the cell concentration at least 2×10 8 / ml.
[0024] Subsequently, the osteosarcoma tibial orthotopic model was established with SPF grade 4-week-old nude mice (BALB / c-nu / nu), a total of 20 mice, 5 mice in each group, 4 groups in total. Animal feeding and manipulation were performed in accordance with the requirements of the Animal Ethics Committee of Shanghai Jiaotong University School of Medicine. Anesthetize with 3.5% sodium pentobarbital, ...
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