摘要
为探索如何快速获取活性菌株的筛选模式 ,组合使用海虾生物致死法和tsFT2 10细胞的流式细胞术筛选模型 ,对从青岛前海和胶州湾潮间带海泥、动植物样品中分离的 162株海洋微生物 ,进行了抗肿瘤活性的分级组合筛选。经海虾生物致死法一级初筛 ,得到活性菌株 5 8株 (LC50 <5 0 0 μg/mL) ,阳性率为 3 5 .8%。对初筛有效的 5 8株菌利用流式细胞术筛选模型进行二级复筛 ,得到对tsFT2 10细胞有各种活性的活性菌株共 2 5株 ,阳性率为复筛菌株数的 43 .1%、初筛菌株总数的 15 .4% ,其中 ,诱发细胞坏死伴有凋亡诱导活性的菌株 7株 (占总菌株的 4.3 % ) ,具有细胞周期抑制活性的菌株 18株 (占总菌株的 11.1% )。该分级组合筛选模式与流式细胞术筛选模型的单独筛选模式相比 ,无漏筛 ,具有成本低、速度快、宜于大规模筛选等特点。
In the present study, a gradated-combinatorial screening for antitumor-active microbial strains was carried out by the gradated combination of a 96-well plate lethality bioassay using brine shrimp and a flow cytometric bioassay using mouse tsFT210 cells. At the first step, 162 strains of marine-derived microbes, isolated from marine samples collected at Qingdao Offing and Jiaozhou Bay, were first screened by Brine Shrimp Lethality assay to obtain 58 strains showing lethiferous activity on brine shrimp, which were 35.8% of the total 162 strains tested. At the second step, 58 strains were subjected to the flow cytometric second screening using mouse tsFT210 cells and a total of 25 strains were found to show antitumor activities on the tsFT210 cells, which accounted for 43.1% of the 58 strains and 15.4% of the total 162 strains, respectively. Among them, 7 trains(4.3% of the total 162 strains) showed mainly necrotic cytotoxicity, accompanying apoptosis-inducing activity, and 18 strains (11.1% of the total) showed call inhibitory activity, which coincided with the results of direct screening of 162 strains by the flow cytometric bioassay using tsFT210 cells. The present study provides a practical gradated-combinatorial screening approach for large-scale speedy-screening of cell cycle inhibitory, apoptosis-inducing and/or cytotoxic microbial strains effective on mammalian cancer cells.
出处
《中国海洋大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第1期38-42,共5页
Periodical of Ocean University of China
基金
国家高技术研究发展计划(2001A624020)
国家杰出青年科学基金(39825126)
国家自然科学基金(30300433)
国家重点基础研究发展规划项目计划(G1998051113)
国家高技术发展计划青年基金(2002AA628130)
山东省科技攻关计划(0121100107)
山东省自然科学基金重点项
关键词
海洋微生物
分级组合筛选
筛选模型
抗肿瘤
tsF210
海虾
细胞周期
细胞凋亡
细胞毒
marine-derived microorganisms
gradated-combinatorial screening
bioassay
antitumor
tsFT210
brine shrimp
cell cycle
apoptosis
cytotoxicity
