Application of mycosubtilin to preparation of anti-tumor drug
The technology of anti-mycosubtilin and anti-tumor drug is applied in the application field of anti-mycosubtilin in the preparation of anti-tumor drugs, and can solve the problems such as the anti-cancer function and application of no-known anti-mycosubtilin, and achieve side effects. small effect
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Embodiment 1
[0033] Example 1: Preparation of Mycosubtilin
[0034] A method for extracting mycosubtilin used in anti-tumor, specifically comprising the following steps:
[0035] (1) Inoculate the preserved Bacillus subtilis into a test tube containing 5 mL of NB culture solution, and incubate at 37 °C, 180 r min -1 Cultivate under shaking for 12 hours to obtain seed solution;
[0036] (2) Inoculate the seed solution obtained in step (1) in a conical flask containing fermentation medium at a volume ratio of 1:10, and -1 Shake and cultivate for 18-36 hours to obtain a fermented liquid, remove bacteria by suction filtration, and collect aseptic fermented liquid.
[0037] (3) Add concentrated hydrochloric acid to the sterile fermentation liquid obtained in step (2), adjust the pH value to 2.0, let it stand at 4°C for 10 hours, and collect the precipitate after centrifugation to obtain a crude extract.
[0038] (4) Fully dissolve the crude extract obtained in step (3) with 20 mL of n-butano...
Embodiment 2
[0041] Example 2: Application of Mycosubtilin in the Preparation of Antitumor Drugs
[0042] In this example, on the basis of Example 1, using in vitro cell culture technology, different doses of mycosubtilin were added to human liver cancer cell Bel7404 adherently cultured in vitro, and mycosubtilin was not added or DDP was blank. As a control, the other conditions were exactly the same, treated for a certain period of time, and the amount of active cells was measured by MTT method to investigate the lethal effect of mycosubtilin on mouse and human liver cancer cells in vitro; After a period of time, the cells were collected and detected by flow cytometry, and quadrant plots were made to investigate the effect of mycosubtilin on inducing the apoptosis of mouse liver cancer cells H22 and human liver cancer cells Bel7404.
[0043] (1) Cell culture
[0044] The cell culture medium was DMEM complete medium containing 8%-15% fetal bovine serum, 5% penicillin-streptomycin double a...
Embodiment 3
[0052] Example 3: Application of Mycosubtilin in the Preparation of Antitumor Drugs
[0053] (1) Effect of mycosubtilin on the growth of liver cancer tumors in mice
[0054] In this embodiment, on the basis of Example 1, 48 subcutaneous tumor-bearing (liver cancer) mice (Kunming mice) were taken, and 12 were divided into 4 groups at random. 1mg / kg mycosubtilin solution, 3mg / kg mycosubtilin solution, and 2mg / kg DDP solution were injected intraperitoneally into the mice, and the four groups were respectively recorded as CK and 1mg / kg J-15 -MYC, 3mg / kg J-15-MYC, 2mg / kg DDP group. Among them, the first three groups were injected once a day, and the 2mg / kg DDP group was injected once every other day. A total of 20 days of drug treatment. Vernier calipers were used to track and measure the tumor volume changes of the mice in each group, and the mice were sacrificed at day 20 of treatment, the tumors were dissected out, the tumor weight was weighed, and the mortality rate of the m...
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