PER (protein efficiency ratio)2 protein agonist polypeptide and application thereof
An agonist and protein technology, applied in the field of PER2 protein agonist peptides, can solve problems such as drugs for esophageal cancer that have not yet been developed, and achieve the effect of inhibiting proliferation and improving survival rate
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Embodiment 1
[0019] Effect of PER2 protein agonist polypeptide on the proliferation of human esophageal cancer cell Eca109.
[0020] The MTT colorimetric method was used. The Eca109 cells with logarithmic growth were added to 96-well culture plate at 1.0×105, and cultured for 24 hours. Different concentrations of experimental drug PER2 protein agonist polypeptide and positive control drug vincristine were added to the experimental wells and positive drug control wells; blank The same volume of solvent was added to each group. Set five duplicate wells in each well, culture for 48h, add MTT to each well at 0h, 2h, 8h, 14h, 20h, 24h, 36h, and 48h respectively, add DMSO after 4h of action, 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 the experimental group / absorbance value of the control group) × 100%. The calculated IC50 of the test drug was 5.01 μM. When the concentra...
Embodiment 2
[0022] Effect of PER2 protein agonist polypeptide on the migration of human esophageal cancer cell Eca109.
[0023] 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; Eca109 cells growing in pairs were added to a 24-well culture plate at 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 PER2 protein agonist polypeptide 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. Take pictures at 0, 6, 12, 2...
Embodiment 3
[0027] The in vivo activity of the PER2 protein agonist polypeptide was tested with a tumor model.
[0028] The Eca109 tumor model was established, and the positive control drug was vincristine; the same volume of solvent was added to the blank group, and 3 doses were set to the experimental group: 4, 8, and 16 mg / Kg. After 21 days, the number of surviving mice was observed, and the survival rate was calculated. The results showed that the PER2 protein agonist polypeptide 2 can effectively protect mice and improve the survival rate of tumor-bearing mice, with the survival rate reaching 70.0%.
[0029]
PUM
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