FOXM1 antagonistic polypeptide as well as derivative and application thereof
A derivative and antagonistic technology, applied in the fields of biotechnology and biomedicine, can solve the problems of anti-tumor drugs on the market, and achieve the effect of inhibiting the proliferation of cancer cells, promoting apoptosis, and huge social and economic benefits.
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Embodiment 1F
[0037] Example 1FIP-1 can be combined with FOXM1 in HepG2 cells
[0038] The HepG2 cells having good state were seeded in a 12-well plate containing the climbing, and fresh medium was added to culture overnight, and the labeled polypeptide, FIP-1 and control group polypeptide TAT were added the next morning, and the culture plate was cultured. The slides that have been climbed in the cells were dipped 3 times with PBS, 3 min each time; 4% polymethyledealdehyde fixed climb for 15 min, PBS dipped 3 times, 3 min each time; 0.5% Triton X-100 (PBS preparation) Room temperature Transparent 20min (antigen expressed in the cell membrane omitted this step); PBS dipped 3 times, 3 min, water absorbing paper to dry PBS, add normal goat blood on a slide, room temperature to close 30 min; The water absorbent is suiled to remove the enclosure, not wash, each slide is dropped with a sufficient amount of diluted one-resistant and placed in a wet cartridge, incubate overnight at 4 ° C; second day p...
Embodiment 2F
[0039] Example 2FIP-1 can inhibit the proliferation of HepG2 cells
[0040] The heporous cell hepg2 was seeded in a 6-well cell culture plate in 100 / well, and the medium per well was 2 mL, and 24 h was cultured. The medium and drugs were replaced every two days, continuously cultured, and the cell group contains more than 50 cells, and 0.1% crystalline purple color, statistically, the number of cloned formations in each group was carried out with methanol fixed experimental group and control group. image 3 It can be seen from the figure that FIP-1 can effectively inhibit the number of cells of the cells, inhibit the formation of clones, confirming that FIP-1 can effectively inhibit cell proliferation.
Embodiment 3
[0041] Example 3CCK-8 detection of inhibitory effects of FIP-1 on HepG2 cells
[0042] In a 10 cm culture medium inoculated in a 10 cm culture medium, it was cultured to 95% by 10% FBS DMEM medium. After the cells reached the desired density, the medium was removed, washed twice with PBS, add 1 ml of trypsin, and incubate 37 degrees. After 1 min, 1 ml of 10% FBS DMEM medium was added, and the dispersion cell was tapped, and 10 μl of cell suspension was added to 1 ml of DMEM medium to prepare cell suspension, and cells prepared by cells were counted. Quantity, then press each hole 5 × 10 3 The cultured in 96-well plates for 24 hours. On the second day, the cell culture medium was abandoned, and fresh medium and different concentrations of FIP-1 were added to the culture plate, and the culture plate was incubated for 48 hours in the incubator, and 10 μl of CCK-8 solution was added to each well (note not to be generated in the well. Bubbles, they affect the OD value of the OD value t...
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