海洋微生物抗肿瘤活性产物的筛选、菌种鉴定及产物的初步分析-食品工程专业论文.docxVIP

上海海洋大学硕士学位论文 上海海洋大学硕士学位论文 II II μg?mL-1；菌株 N16 代谢产物对 MCF-7 和 BEL-7402 两种肿瘤细胞的 IC50 值分别为 84 μg?mL-1 和 133 μg?mL-1。通过对菌株 S-1 和 N16 的 16S rDNA 序列测定、同源 性比对分析及生理生化特征研究，确定菌株 S-1 为短芽孢杆菌属（Brevibacillus sp.）， N16 为芽孢杆菌属（Bacillus sp.）。 2、采用 MTT 活性追踪法，通过水饱和正丁醇萃取法、CM Sepharose Fast Flow 阳离子交换色谱法和制备型 C18 HPLC 反相色谱法等分离手段进一步从菌株 S-1 代谢产物中分离纯化出一种能抑制肿瘤细胞增殖的多肽，命名为 SBP（Peptide from Brevibacillus sp. S-1，SBP）；采用质谱技术对 SBP 的分子量进行测定，其分子量为 1570 Da。 3、采用 MTT 法测定了抗肿瘤多肽 SBP 对人肝癌细胞 BEL-7402 的抑制作用， 结果显示：阳性对照丝裂霉素对人肝癌细胞 BEL-7402 的 IC50 值为 1.44 μM，SBP 对人肝癌细胞 BEL-7402 的 IC50 值为 7.15 μM；通过倒置相差显微镜观察活性多肽 SBP 作用后肝癌细胞 BEL-7402 的细胞形态。结果显示：SBP 能够使细胞表面粗糙、 细胞体积变小、慢慢变圆、细胞间连接变少，最终导致细胞大部分脱落，贴壁细 胞进一步减少，细胞皱缩成一个圆点，细胞间连接几乎全部消失。 初步研究结果显示：抗肿瘤多肽 SBP 可以改变人肝癌细胞 BEL-7402 的细胞 形态，能抑制肝癌细胞 BEL-7402 的增殖，有望成为抗肿瘤药物的候选化合物。 关键词：海洋微生物；筛选；鉴定；抗肿瘤；纯化 III III Screening of antitumor metabolites from marine microorgan- isms, strain identification and primary investigation of the metabolites ABSTRACT According to the statistics from World Health Organization (WHO), malignant cancer is still the most serious disease threating human healt h. Excepting inhibiting the growth of cancer cells, most of current antitumor agents act against normal mammalian cells, consequently causing severe side e ffects. So it becomes an important assignment to find a kind of anti-tumor drugs with efficiency and low side effects. Finding and re- searching high-efficiency antineoplastic agent from the nature is a lways a hot spot in the field of oncotherapy. Microbes and their metabolites, because of their diversity, were overlooked by experts and scholars at whole world for a long time. But the traditional drugs from the land were limited, and another huge reservoir, marine organisms, had been ignored for a long time. Many inspiring achievement had been made by scie ntist on the finding of marine compounds with anti-tumor activity. Marine organisms repre- sented an essentially unexploited reservoir for products with potentially biological and pharmacological