Medicine for treating malignant cerebroma and breast cancer by combining gene and chemotherapy medicine and preparation method thereof
A technology of combined chemotherapy and gene therapy, applied in the field of biomedicine, can solve the problems of low water solubility and limit the clinical application of anti-cancer, and achieve the effect of inhibiting tumor growth, reducing cell migration ability, and reducing cell survival rate.
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Embodiment 1
[0033] (1) Dissolve 2 mg of polyamidoamine dendrimer in 1 mL of buffer solution to obtain solution A. (2) Dissolve 0.1 mg of oligonucleotide in 1 mL of buffer solution to obtain solution B. (3) Dissolve 0.05 mg of paclitaxel in 1 mL of buffer solution to obtain solution C. At room temperature, mix 10 parts by weight of solution A with 1 part by weight of solution B, incubate for 20 minutes, and then add 5 parts by weight of solution C to obtain antisense microRNA-21 gene therapy combined with paclitaxel chemotherapy A new type of malignant brain tumor
[0034] treatment system. Its transmission electron microscope pictures are as figure 1 .
[0035] List of specific production conditions:
[0036]
Embodiment 2
[0038] Inoculate U251 human glioma cells and MCF-7 breast cancer cells in 6-well culture dishes respectively, and culture the cells to 80% coverage. In the serum-free state, with PAMAM as the carrier, FITC-labeled antisense microRNA-21 was transfected into U251 cells and MCF-7 cells respectively. The final mass concentration of antisense microRNA-21 was 5 mg / L. The amino group of PAMAM The ratio of phosphate to antisense microRNA-21 is 16:1. The transfection rate was detected by flow cytometry, as shown in Figure 2. Figure (a) is the transfection efficiency of PAMAM after polyamide and antisense microRNA-21 are compounded, and figure (b) is the transfection efficiency of PAMAM after polyamide is compounded with antisense microRNA-21 and paclitaxel;
Embodiment 3
[0040] In place of 1 x 10 4 / ml U251 human glioma cells and MCF-7 breast cancer cells were seeded in 96-well cell culture plates, 500 μl per well, placed at 37°C, 5% CO 2 Incubator for 24h. After the cells adhered to the wall, 8 paclitaxel single-drug control groups with different concentrations were added, and antisense microRNA-21+paclitaxel was the combined drug group (0.025, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 1.0 μmol / L ). Transfection of oligonucleotides was carried out according to the instructions, and the final mass concentration of antisense microRNA-21 oligonucleotides was 5 mg / L. Each concentration combination was repeated for 6 wells. After 48 hours of transfection, 20 μl of MTT (5 mg / mL) was added to each well and incubated at 37° C. for 4 hours. Discard the medium, add 150 μl dimethyl sulfoxide (DMSO) and shake to dissolve for 15 minutes, measure the absorbance (A) at 570 nm with a microplate reader, and measure the survival rate of the cells according to the abso...
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