【作者】 何平平；

【导师】 赵敏；

【作者基本信息】 东北林业大学 ， 微生物学， 2008， 硕士

【摘要】 本文研究了粘质沙雷氏菌中灵杆菌素的提取方法,采用热酚水法提取灵杆菌素以及对提取出的脂多糖粗制品进行了纯化,并通过MTT法、电镜、PT-PCR和DNALadder、流式等方法对灵杆菌素的抗肿瘤增殖作用进行初步研究。发现灵杆菌素对结肠癌LS174t细胞有明显的抑制作用并呈浓度时间依赖关系;其诱导LS174t细胞出现典型的细胞凋亡超微结构改变;RT-PCR检测发现经不同浓度(5、10、25、50、100μg·ml^-1)的灵杆菌素处理后细胞中bcl-2的表达下调,bax的表达上调;DNA电泳出现凋亡特有的“梯子”条带;流式细胞仪检测发现灵杆菌素可引起LS174t细胞凋亡并阻滞于S期。表明灵杆菌素能有效地通过诱导凋亡抑制LS174t细胞生长,其机制可能与抑制抗凋亡因子bcl-2的表达,增强凋亡因子bax的表达有关。更多还原

【Abstract】 This paper studies on the prodigiosin was prepared from Serratia marcescens with hot phenol water method and purified by ultracentrifugafion.Preliminary study by MTT assay,electron microscopy,PT-PCR,DNA Ladder and flow cytometfic inhibited tumor cell proliferation with prodigiosin.Prodigiosin inhibited growth and proliferation of colon cancer cells LS174t in dose and time dependent manner.Prodigiosin induced colon cancer cells LS174t to undergo typically apoptotic.RT-PCR showed that after treatment with prodigiosin for different concentrations(5、10、25、50、100μg.ml^-1),expression lever of Bcl-2 was reduced,Bax was increased.Agarose gel eletrophoresis of DNA revealed the apoptosis characteristic.Prodigiosin arrested LS174t cells in S-phase.The results showed that prodigiosin can induce the apoptosis of LS174t and may be mediated by down-regulation of Bcl-2 and up-regulation of Bax.更多还原