海洋放线菌和真菌野生株及其突变株的分离培养与抗肿瘤活性筛选  

Isolation of wild and mutant strains of marine Actinomyces and fungi and screening for their antitumor activity

作　　者：孙玉雯[1] 崔承彬[1] 韩小贤[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《军事医学科学院院刊》2008年第6期532-536,共5页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金(30572279);国家高技术研究发展计划("863"计划)(2007AA09Z411)

摘　　要：目的:从海泥样品中分离培养并筛选获取野生型与突变型药源微生物抗肿瘤活性菌株。方法:通过单菌落挑选与划线培养,分离纯化野生菌株;利用核糖体工程抗性筛选技术,筛选获得抗生素抗性突变株;用K562细胞,采用MTT法测试发酵样品的抗肿瘤活性。结果:从天津塘沽驴驹河渤海湾潮间带海泥样品中分离得到放线菌126株、真菌29株,其中,在100μg/ml样品浓度下抑制率大于20%的放线菌17株、真菌3株,在1000μg/ml样品浓度下抑制率小于5.5%的无活性放线菌19株。以1000μg/ml样品浓度下抑制率为4.8%的无活性放线菌野生株L8-5x为原始菌,经抗性筛选获得链霉素抗性突变株78株,其中在100μg/ml样品浓度下抑制率大于25%的活性突变株2株。结论:所获野生型与突变型抗肿瘤活性菌株为后续新药筛选提供了药源微生物活性菌株。由无活性菌株转化获取活性突变株的研究结果表明,已分离得到的大量无活性菌株也是可转化获取药源活性突变株的很好资源。Objective：To isolate and screen the wild and mutant strains with antitumor activity from sea soil samples. Methods： The wild strains of marine microbes were obtained by single colony isolation and the antibiotic resistant mutant strains were cultured by ribosome engineering resistance screening technique. The antitumor activity was assayed by the MTT method using K562 cells for both the wild and mutant strains with aqueous acetone extracts of the whole broth from their fermentation samples. Results： The 155 wild strains including 126 actinomycetie strains and 29 fungal strains were isolated from the soil samples collected at the tideland of Bohai Bay around Ltijtihe in Tanggu District of Tianjin, China. Among them, samples from 17 actinomycetic and 3 fungal strains showed antitumor activity with the inhibition rates over 20% at the concentration of 100 μg/ml, while samples from 19 actinomyeetic strains showed no inhibitory activity （inhibition rate 〈5.5% ） even at the concentration of 1 000 μg/ml. An actinomyeetic wild strain L8-Sx, the sample from which showed no inhibitory effect on K562 cells with the 4.8% of inhibition rate at 1 000 μg/ml, was used as the initial microbe for the screening of its streptomycin-resistant mutants and 78 mutant strains were obtained. Among the mutant strains obtained, samples from two strains, 2ess4-7 and 2css6-11 resistant to 4 μg/ml and 6 μg/ml streptomycin, respectively, showed antitumor activity with the inhibition rates over 25% at the concentration of 100 μg/ml. Conclusion： Both the obtained wild and mutant strains that produce metabolites showing antitumor activity have provided the microbial strains for further investigation on the bioactive metabolites. Autogenic transformation of L8-Sx to two mutant strains that produce metabolites possessing antitumor activity has suggested that even the microbial strains that do not produce bioactive metabolites might also become an initial strain resource for obtaining the bioactive metabolites-produeing mutant str

关 键 词：海洋微生物 微生物 放线菌 真菌 分离培养 链霉素抗性 抗肿瘤活性 

分 类 号：R91[医药卫生—药学]