ATF 3 (Activating Transcription Factor 3) protein excited polypeptide and application thereof
An ATF3, protein technology, applied in the field of medicine, can solve problems such as increasing cancer risk, and achieve the effect of inhibiting tumor growth, improving survival rate, and promoting expression
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Embodiment 1
[0015] A tumor model was used to detect the anti-tumor effect of ATF3 protein agonist polypeptide in vivo. 6-8-year-old female C57BL / 6 nude mice were randomly divided into 4 groups, 10 in each group. (1) blank group; (2) low-dose ATF3 protein agonist polypeptide group; (3) medium-dose ATF3 protein agonist polypeptide group; (4) high-dose ATF3 protein agonist polypeptide group. To establish ovarian cancer tumor model, the scheme is: add the same volume of solvent to the blank group, and set 3 doses of ATF3 protein activating polypeptide (synthesized by Shanghai Sangong Jill) in the experimental group: 2.5, 5.0, 10.0 mg / Kg, and inject it at multiple points around the tumor . After 21 days of continuous administration, the number of surviving nude mice was observed, and the survival rate was calculated. The results show that the ATF3 protein activating polypeptide can effectively protect nude mice and improve the survival rate of tumor-bearing nude mice, and the survival rate r...
Embodiment 2
[0017] Effect of ATF3 protein agonist polypeptide on ATF3 protein expression in human ovarian cancer cell OVCAR-3T: OVCAR-3T cells were cultured with RPMI-1640 medium containing 10% fetal bovine serum (FBS) at 37°C and 5% CO2 When cultivating in the box to a confluence of more than 90%, digest the cells with 0.25% trypsin, count, and centrifuge at 3000rpm for 5min to collect the cells; add RPMI 1640 culture medium (5ml) with 10% calf serum to suspend the cells, count the cells, and adjust the cells The concentration is 5*10 6 cells / ml and cultured in 6-well culture plates. In the experiment, a blank control group and ATF3 protein agonist peptide groups (5, 10, 20 mg / ml) were set up. After adding 600 μl / well of OVCAR-3T cell suspension to each group, 600 μl of 1640 culture medium was added to the blank control group, and different concentrations of polypeptides were added to each dose group of ATF3 protein agonistic polypeptide. Press 20μl / 1×10 5 Add protein extraction solut...
Embodiment 3
[0020] Proliferation of human ovarian cancer cell OVCAR-3T: Add OVCAR-3T cells to RPMI1640 culture medium (5ml) with 10% calf serum to suspend cells, count the cells, and adjust the cell concentration to 5*10 6 cells / ml and cultured in 96-well culture plates. In the experiment, a negative control group and ATF3 protein agonist peptide groups (5, 10, 20 mg / ml) were set up. After adding 100 μl / well of OVCAR-3T cell suspension to each group, 100 μl of 1640 culture medium was added to the negative control group, and different concentrations of polypeptides were added to each dose group of ATF3 protein agonistic polypeptide. 37°C cell culture incubator for static culture for 48 hours, after the end of the culture, add 20 μl MTT to each well, continue to cultivate for 4 hours, finally discard all the solution in each well, add 100 μl DMSO to each well, shake, and use a microplate reader to detect the OD value at 570nm. 5 in parallel.
[0021] Table 1 Proliferation effect of ATF3 p...
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