Application of mauremys mutica polypeptide mixture and antitumor drug
An anti-tumor drug, the technology of the stone turtle, applied in the field of anti-tumor drugs and drugs, can solve the problems of poor sensitivity of external growth inhibitory signals, multi-drug resistance of tumors, and it is difficult for anti-tumor drugs to kill tumor cells. Good application prospect, the effect of inhibiting proliferation
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Embodiment 1
[0049] Embodiment 1, the preparation of stone money turtle polypeptide mixture
[0050] Mix 10 g of minced meat of the stone turtle with 80 g of ultrapure water, add neutral protease (25000 U, 37313.43 U / g) under the conditions of temperature of 45 ° C and pH of 7.5 and fully react for 10 h to obtain a reaction solution.
[0051] Inactivate the reaction solution at 85°C for 10 minutes, centrifuge at 8000r / min for 30 minutes, remove the precipitate and retain the supernatant, freeze-dry the supernatant to obtain the crude product, that is, the stone money turtle polypeptide mixture
[0052] The obtained stone money turtle polypeptide (GDP) mixture was diluted to an original concentration of 1 g / mL, and stored at 4°C for future use.
Embodiment 2
[0053] Embodiment 2, the determination of the antitumor performance of the stone money turtle polypeptide mixture
[0054] 1. Determination of inhibition of tumor cell proliferation by stone money turtle polypeptide
[0055] Take the tumor cell lines in the logarithmic growth phase to make a certain concentration of cell suspension, inoculate into 96-well plate, inoculate 5000 cells (100 μL) in each well, and place in 5% CO 2 , in a 37°C incubator, discard the original medium after culturing for 24 hours, wash it once with PBS, and then add different concentrations of the acronym tortoise peptide mixture solution, set 3 parallel wells for each concentration, and set up a control group with PBS at the same time , set 5% CO 2 , in a 37°C incubator, after incubation for 24 hours, discard the original medium, add PBS to wash 2 times, then add 10 μL MTT solution to each well, place in the incubator and incubate for 4 hours, absorb the MTT solution, and then add 100 μL DMSO solutio...
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