两株海洋微生物抗肿瘤代谢产物的筛选及其菌种鉴定  

Screening of Antitumor Metabolites From Two Marine Microorganisms and Their Strain Identification

作　　者：衣尧[1,2] 郑兰红[1] 陈世建[1] 盛军[1] 孙谧[1] 

机构地区：[1]农业部海洋渔业可持续发展重点实验室中国水产科学研究院黄海水产研究所,青岛266071 [2]上海海洋大学食品学院,上海201306

出　　处：《渔业科学进展》2014年第6期114-119,共6页Progress in Fishery Sciences

基　　金：中国水产科学研究院基本科研业务费(2014B01YQ01);山东省科技发展计划项目(2014GSF121016);中央级公益性科研院所基本业务费专项资金项目(206030220130017;20603022012013)共同资助

摘　　要：对5株海洋微生物进行发酵培养,发酵液离心后,收集上清液,采用饱和正丁醇萃取制备海洋微生物的代谢产物,采用MTT活性跟踪法,以BEL-7402、RKO、A549、U251和MCF-7等细胞系为模型,对代谢产物进行抗肿瘤活性的筛选。结果显示,菌株S-1和N16代谢产物的活性组分具有较好的抗肿瘤活性:菌株S-1对MCF-7、U251和BEL-7402三种肿瘤细胞的IC50值分别为44、102和82μg/ml;菌株N16对MCF-7和BEL-7402两种肿瘤细胞的IC50值分别为84、133μg/ml。对菌株S-1和N16进行了16S r RNA序列分析、生理生化特征鉴定,结果显示,菌株S-1为短芽孢杆菌属(Brevibacillus sp.),N16为芽孢杆菌属(Bacillus sp.)。通过显微镜观察了菌株Brevibacillus sp.S-1和Bacillus sp.N16代谢产物的活性组分对BEL-7402细胞形态的影响,表明两株菌的活性组分可以改变肝癌细胞BEL-7402的细胞形态,抑制细胞增殖。该研究为新型抗肿瘤先导化合物的发现提供了新的海洋微生物种质资源。Marine microorganisms are the important resources in the natural products with biological activity. Finding antitumor drugs and compounds from germplasm resources has become a hot topic. The current study investigated the biological role of the fermented products of marine bacteria isolated from marine strains in several cancer cell lines. Five marine strains were fermented, and the supernatant was collected after centrifugation as total products that were fractionated with n-butyl alcohol. The pooled organic layer was evaporated using a rotary evaporator under reduced pressure, and the peptide pellet was dissolved in phosphate buffer for further usages. Antitumor metabolites of five marine microorganisms were screened by MTT method in several cancer cell lines. The result showed that the fermented products of strain S-1 and N16 inhibited cancer cell proliferation. Specifically, fermented product from strain S-1 attenuated MCF-7, U251 and BEL-7402 cell proliferation and the IC50 were 44μg/ml, 82μg/ml and 102μg/ml, respectively. Moreover, fermented product from strain N16 inhibited MCF-7 and BEL-7402 cell proliferation and their IC50 was 84μg/ml and 133μg/ml, respectively. The fermented products of S-1 and N16 induced the morphological change of the BEL-7402 cells. According to their 16S rRNA sequences and physiological biochemical properties,strains S-1 and N16 were identified as Brevibacillus sp. and Bacillus sp., respectively. The further research is to find new antitumor compounds from the fermented products of marine bacteria Brevibacillus sp. S-1 and Bacillus sp. N16. This study supports that the germplasm resources of marine microorganisms may be a novel resource for the discovery of novel anti-tumor compounds.

关 键 词：海洋微生物 菌种鉴定 代谢产物 抗肿瘤活性 

分 类 号：Q93[生物学—微生物学]