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目的:研究鱿鱼墨多肽酸法提取工艺及不同浓度鱿鱼墨多肽对DU-145和PC-3细胞的增殖抑制和诱导凋亡作用。方法:以料液比、加酸量、酸解时间和酸解温度为因素,以酸解液的氨基氮含量为指标,通过正交试验确定提取鱿鱼墨多肽的酸解条件。采用CCK-8法检测其对DU-145和PC-3的生长抑制作用。采用Annexin V-FITC/PI双标记流式细胞术检测和AO/EB双染法,观察鱿鱼墨多肽对DU-145和PC-3细胞的早期凋亡情况。结果:酸法提取鱿鱼墨多肽最佳组合为0.02 mol/L盐酸、酸提料液比1∶1、酸解时间2 h、酸解温度35℃;鱿鱼墨多肽对DU-145和PC-3细胞有增殖抑制作用,能诱导DU-145和PC-3细胞凋亡。结论:鱿鱼墨多肽能抑制DU-145和PC-3细胞增殖,并诱导其细胞凋亡。
OBJECTIVE: To study the acid extraction process of squid ink polypeptide and the proliferation inhibition and apoptosis induction of DU-145 and PC-3 cells with different concentrations of squid ink peptides. Methods: Taking the ratio of solid to liquid, the amount of acid, the time of acid hydrolysis and the temperature of acid hydrolysis as factors, the content of amino nitrogen in acid solution was taken as an index to determine the optimal conditions for the extraction of squid ink polypeptide by orthogonal test. The growth inhibition of DU-145 and PC-3 was tested by CCK-8 assay. Annexin V-FITC / PI double-labeled flow cytometry and AO / EB double staining were used to observe the early apoptosis of DU-145 and PC-3 cells. Results: The optimal combination of squid ink for acid extraction was 0.02 mol / L hydrochloric acid, the ratio of acid to material was 1: 1, the acid hydrolysis time was 2 h and the acid hydrolysis temperature was 35 ℃. Cell proliferation inhibition, can induce DU-145 and PC-3 cell apoptosis. Conclusion: Squid ink polypeptide can inhibit the proliferation and induce the apoptosis of DU-145 and PC-3 cells.