An enzymolysis polypeptide and its use for inhibiting the invasion and metastasis of small cell lung cancer stem cells
A technology of lung cancer stem cells and small cell lung cancer, applied in the field of medicine, to achieve the effect of preventing tumor metastasis
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Embodiment 1
[0027] The preparation of embodiment 1 enzymolysis polypeptide
[0028] 1. Experimental materials
[0029] 1. Instrument reagents
[0030] The ultrafiltration cup and ultrafiltration membrane were purchased from Shanghai Mosu Scientific Equipment Co., Ltd.;
[0031] Alkaline protease 37017 (1:2.5U / g) was purchased from Novozymes.
[0032] 2. Experimental samples
[0033] Fresh cicada monkeys just unearthed from trees were collected, stored in water, and frozen at -20°C. Thaw naturally at room temperature before use.
[0034] 2. Experimental method
[0035] 1. Enzymatic hydrolysis method
[0036] Wash the cicada monkeys and grind them into minced meat. Take 10g of the minced meat and put it in 50mL of ultrapure water, add alkaline protease for enzymolysis. The amount of alkaline protease added is 0.45% of the weight of the substrate minced meat. The enzymatic hydrolysis conditions are: pH value, 8.5; enzymatic hydrolysis temperature, 55°C; enzymatic hydrolysis time, 10 h...
Embodiment 2
[0049] Example 2 Effects of Enzymatically Hydrolyzed Polypeptides on the Invasion and Metastasis of NCI-H446 Cell Line Lung Cancer Stem Cells
[0050] The experimental method of this example is operated with reference to the literature method (Chen Gongrang et al., Vitexin Inhibition of Small Cell Lung Cancer NCI-H446 Cell Line Lung Cancer Stem-Like Cell Invasion and Metastasis, Zhongnan Pharmacy, Volume 12, Issue 10, October 2014), And use the vitexin disclosed in this document as positive drug.
[0051] 1. Experimental method
[0052] 1. Culture and passage of human small cell lung cancer NCI-H446 cell line
[0053]Human small cell lung cancer NCI-H446 cell line was purchased from Shanghai Aolu Biotechnology Co., Ltd. Use high-glucose DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 U / mL streptomycin at 37°C, 5% CO 2 cultured under saturated humidity conditions. The cells grow adherently, covering 90% of the bottom of the bottle in about 1-2 day...
Embodiment 3
[0085] Example 3 Safety Evaluation of Enzymatically Hydrolyzed Polypeptides
[0086] Kunming mice, weighing 25-31g, were randomly divided into groups, and the toxicity comparison experiment was carried out by injecting or not injecting the enzymatic polypeptide through the tail vein. The results showed that the enzymatic polypeptide 3 and the enzymatic polypeptide 4 were continuously injected for 45 days at a dose of up to 600mg / kg, without any toxic and side effects, and the anatomical observation of the heart, liver, kidney, and lung showed no abnormalities, and the body weight all increased. The experimental results are shown in Table 3.
[0087] Table 3 Enzymolysis polypeptide toxicity test
[0088]
[0089] The above examples prove that the enzymatic polypeptide provided by the present invention can effectively inhibit the invasion and metastasis of lung cancer stem cell-like cells of the small cell lung cancer NCI-H446 cell line. Toxicology experiments have proved th...
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