Application of interferon-induced protein 6 and medicine
An interferon induction and drug technology, applied in the biological field, to enhance the anti-apoptosis and cell proliferation ability, and reduce the effect of skin radiation damage
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Embodiment 1
[0031] Example 1 Construction of IFI6 overexpression lentivirus and expression verification after infection
[0032]In this example, the whole gene synthesis method (Hanbio Shanghai) was used to synthesize the human IFI6 coding region gene (the nucleic acid sequence is shown in Table 1 below), and the IFI6 overexpression lentivirus LV-IFI6 (target lentivirus) was obtained by packaging with a three-plasmid virus packaging system. and LV-NC (control lentivirus) recombinant lentivirus.
[0033] Table 1 The sequences of primers and genes
[0034]
[0035]
[0036] Human skin keratinocytes HaCaT and human skin fibroblasts WS1 in exponential growth phase were mixed with 10.0×10 4 The density of each well was inoculated in a 6-well plate. After the cells adhered to the wall, the MOI value of the virus provided by the instructions was calculated. Add an appropriate amount of IFI6 overexpression lentivirus or control lentivirus (HBLV-NC) to each well, and incubate at 37°C. , 5%...
Embodiment 2
[0037] Example 2 The effect of IFI6 overexpression lentivirus on the proliferation ability of skin cells after irradiation
[0038] Human skin cells stably overexpressing IFI6 in the exponential growth phase and their control cells were inoculated in 96-well plates, and 4 parallel samples were set up in each group. After the cells adhered to the wall, an absorbed dose of 20Gy was given to HaCaT and WS1 cells respectively. and 5Gy X-ray irradiation. 24 hours after exposure, the cells were processed according to the instructions of the Cell-Light Edu Fluorescence Staining Kit provided by Guangzhou Ruibo Company. After the processing was completed, photos were taken with a fluorescence microscope, and the data were counted and processed. The experiment was repeated three times.
[0039] Such as figure 2 As shown (P<0.01), the results showed that IFI6 overexpression lentivirus can significantly promote the proliferation ability of skin cells after irradiation with ionizing radi...
Embodiment 3
[0040] Example 3 The effect of IFI6 overexpression lentivirus on the apoptosis level of skin cells after irradiation
[0041] Human skin HaCaT, WS1 cells and their control cells stably overexpressing IFI6 in the logarithmic growth phase were seeded in 6-well plates, and 20Gy and 5Gy X-rays were irradiated after the cells adhered to the wall, and the cells were collected 48 hours after irradiation, and used BD's Annexin V / 7AAD PE apoptosis detection kit was used to detect cell apoptosis in each group, and three replicate wells were set up in each group.
[0042] Such as image 3 As shown (P<0.01), the results showed that LV-IFI6 did not affect the apoptosis rate of the 0Gy group; after irradiation, compared with the control lentivirus LV-NC group, the apoptosis rate of the IFI6 overexpression group was significantly reduce.
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