A kind of st2 protein inhibitor polypeptide and its application
A protein inhibitor, 1.ST2 technology, applied in the direction of peptide/protein components, peptides, medical preparations containing active ingredients, etc., can solve problems such as drugs for uterine cancer that have not yet been developed, and achieve the effect of increasing survival rate
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Embodiment 1
[0017] The effect of ST2 protein inhibitor polypeptide on the migration of human uterine cancer cell Hela.
[0018] 10 mg / ml Matrigel (BD Company, USA) was diluted 1:2 with HELA special medium, spread on the transwell chamber membrane, and air-dried at room temperature. HELA cells cultured to the logarithmic growth phase were digested with trypsin digestion solution, collected, washed twice with PBS, and then resuspended with blank HELAs-specific medium. Count under the microscope and adjust the cell concentration to 1 x 10 5 pieces / ml. The test solution of each group was prepared and diluted to 100 μl with blank HELAs-specific medium. The cells were inoculated into the transwell chamber, 100 μl per well, and each group of test liquid was added into the chamber. Add 0.6ml of endothelial cell culture medium containing 5% fetal bovine serum and 1% ECGS to the 24-well plate to stimulate cell migration, in 5% CO 2 , and cultivated at 37°C for 24h. Discard the culture medium i...
Embodiment 2
[0026] The effect of ST2 protein inhibitor polypeptide on the proliferation of human uterine cancer cell Hela.
[0027] The MTT colorimetric method was used. The logarithmic growth of Hela cells was added to 96-well culture plate at 1.0×105 and cultured for 24 hours. Different concentrations of experimental drug ST2 protein inhibitor polypeptide 4 and positive control drug vincristine were added to the experimental wells and positive drug control wells respectively; The same volume of solvent was added to the blank group. Set up five duplicate wells per well, culture for 48h, add MTT to each well at 0h, 2h, 8h, 14h, 20h, 24h, 36h, and 48h respectively, after 4h of action, add DMSO, incubate for 30min, and measure at 620nm in a microplate reader Absorbance A value, according to the formula tumor cell growth inhibition rate=(1-absorbance value of experimental group / absorbance value of control group)×100%. The calculated IC50 of the experimental drug was 7.61 μM. When the conc...
Embodiment 3
[0029] Inhibitory test of ST2 protein polypeptide on the growth of human uterine cancer cell Hela C57BL / 6 transplanted tumor in black mice
[0030] The tumor tissues in the vigorous growth period were taken and ground under sterile conditions, and prepared into 1×10 7 Cell suspension per ml was inoculated subcutaneously in the right axillary of mice with 0.1 ml. Use a vernier caliper to measure the diameter of the transplanted tumor in mice, and wait until the tumor grows to 100-200mm 3 Animals were then randomly grouped. By using the method of measuring the diameter of the tumor, the inhibitory effect of the ST2 protein polypeptide on the tumor of the test animal is dynamically observed. Tumor diameter was measured once every 2 days, and the weight of the mouse was also weighed at the same time as each measurement. The experimental group was subcutaneously injected with polypeptide in the left armpit, while the negative group was given the same amount of normal saline for ...
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