Cyclin D protein inhibitor polypeptide and application thereof
A technology of protein inhibitors and injections, which is applied in the field of polypeptides for the treatment of mantle cell lymphoma, can solve problems that have not yet been developed, and achieve the effect of increasing survival rate and inhibiting tumor growth
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Embodiment 1
[0018] Effect of Cyclin D protein inhibitor peptide 1 on migration of Mino in human mantle cell lymphoma cells.
[0019] Using scratch test. First use a marker pen on the back of the 24-well plate, and use a ruler to draw horizontal lines evenly, about every 0.5~1cm, across the holes. Three lines were passed through each well; Mino cells growing in pairs were added to a 24-well culture plate at a density of 1.0×105, and cultured for 24 hours. On the second day, compare the ruler with a 10 μl pipette tip, scratch perpendicular to the horizontal line on the back, and use the intersection of the scratch and the horizontal line on the back as a fixed observation point; add different concentrations of experimental wells and positive drug control wells respectively. The drug Cyclin D protein inhibitor polypeptide 1 and the positive control drug vincristine; the same volume of solvent was added to the blank group, and five replicate holes were set in each well; 2 Incubator, culture...
Embodiment 2
[0023] Effect of Cyclin D protein inhibitor peptide 1 on the proliferation of human mantle cell lymphoma cells Mino.
[0024] The MTT assay was used to detect the activity of the peptides in inhibiting the growth of Mino cells. Mino cells at 37°C, 5% CO 2 When the incubator was cultivated to a confluence of more than 90%, it was digested with trypsin and collected, and the cells were resuspended in the culture medium and counted under the microscope, and the cell concentration was adjusted to 2×10 4 cells / ml, inoculate the cell suspension into 96-well plate, 100 μl / well, and incubate at 37°C, 5% CO 2 Incubate overnight in the incubator. The polypeptides are diluted with culture medium to each predetermined concentration. Endostar was diluted with culture medium to the final concentration. After the cells were completely adhered to the wall, each dilution was added to a 96-well plate (100 μl / well). Adding the integrin blocking agent polypeptide dilution was used as the adm...
Embodiment 3
[0031] Inhibition of Cyclin D protein inhibitor polypeptide 1 on human mantle cell lymphoma cell Mino xenograft tumor growth in nude mice
[0032] Take the human mantle cell lymphoma Mino cell line in the logarithmic growth phase, and prepare 5 × 10 cells under sterile conditions. 7 / ml cell suspension, inoculated subcutaneously in the right armpit of nude mice with 0.1ml. Use a vernier caliper to measure the diameter of the transplanted tumor in nude mice, and wait until the tumor grows to 100-200mm 3 Animals were then randomly grouped. Using the method of measuring tumor diameter, dynamically observe the anti-tumor effect of the tested polypeptide. Tumor diameter was measured every 2 days. The method of administration was by tail vein injection. The negative control group was injected with the same amount of normal saline once a day; the paclitaxel group was given 10 mg / kg once a week; the endostar group was given 2.5 mg / kg once a day; 10mg / kg, 5mg / kg, administered once...
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