Application of adenovirus mediated interleukin-24 for preparing radiotherapy hypersitization medicine
A technology of interleukin and adenovirus, applied in drug combination, gene therapy, antineoplastic drugs, etc., can solve the problem that Ad-IL-24 radiosensitizer has not been reported.
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Embodiment 1
[0031] (1) Cell culture: SPC-A1 cells use RPMI1640 complete medium (10% FCS), at 37 ° C, 5% CO 2 under cultivation. Subculture once every 2 to 3 days.
[0032] (2) Determination of amplification and titer of AdV, Ad-IL-24 adenovirus
[0033] Infect 70% of the adherent QBI-293A cells with AdV and Ad-IL-24, collect the cells after 48 hours, freeze and thaw the cell suspension at -80°C and 37°C four times, take the supernatant, and repeatedly amplify, the virus Store at -80°C.
[0034] Culture and grow QBI-293A cells in good condition, digest with trypsin, count the cells, and dilute the cell concentration to 10 5 cells / ml, inoculate the cells at 100 μl per well on a 96-well plate, and after culturing for 24 h, the harvested recombinant virion was injected at 10 -4 、10 -5 、10 -6 、10 -7 、10 -8 After dilution, inoculate 1 row at 100 μl per well for each dilution, at 37°C, 5% CO 2 After culturing in the cell incubator for 18 hours, the fluorescence was counted under a fluor...
Embodiment 2
[0065] Example 2, the tumor inhibitory effect of Ad-IL-24 and combined radiotherapy on the transplanted tumor of SPC-A1 lung adenocarcinoma in nude mice.
[0066] (1) Establishment of SPC-A1 human lung adenocarcinoma tumor-bearing nude mouse model
[0067] SPC-A1 human lung adenocarcinoma cells were completely cultured with RPMI1640 complete medium (10% FCS) in 5% CO2, and conventionally cultured at 37°C. The SPC-A1 human lung cancer cells in the logarithmic growth phase were washed with PBS, digested with 0.25% trypsin, stopped with serum-free medium, centrifuged at 1000r / min for 5min to harvest the cells, and adjusted the cell concentration after counting the cells. 3×10 7 / ml of cell suspension. Twenty-five 4-week-old SPF grade BALB / c male nude mice were routinely disinfected with Aneriodine in the right anterior armpit of the nude mice, and each nude mouse was subcutaneously injected with a cell suspension of 3.0×10 6 / 100μl, and observe the growth and tumor formation o...
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