粘细菌<em>Myxococcus macrosporus</em> STXZ54抗肿瘤活性物质的分离制备及其活性测定

doi:10.13523/j.cb.20140910

中国生物工程杂志

2014, Vol. 34

Issue (9): 63-71 DOI: 10.13523/j.cb.20140910

技术与方法

粘细菌Myxococcus macrosporus STXZ54抗肿瘤活性物质的分离制备及其活性测定

文也, 唐少军, 肖蓉, 丁学知, 黄同龙, 雷良欢, 夏立秋

湖南师范大学生命科学学院微生物分子生物学国家重点实验室培育基地长沙 410081

Separation and Evaluation of Anticancer Components from Myxococcus Macrosporus STXZ54

WEN Ye, TANG Shao-jun, XIAO Rong, HUANG Tong-long, LEI Liang-huan, DING Xue-zhi, XIA Li-qiu

State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, Province 410081, China

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摘要：

粘细菌作为第三大类次级代谢产物产生菌，是挖掘抗肿瘤新药物的重要资源。将本实验室保藏的粘细菌菌株Myxococcus macrosporus STXZ54进行发酵培养后，通过对发酵液进行硫酸铵沉淀、丙酮沉淀等，分析了该菌株发酵液中抗肿瘤活性物质的性质，采用高效液相色谱技术对发酵液中的抗肿瘤活性物质进行了分离，并对该物质进行了肿瘤细胞毒性的测定，利用激光共聚焦显微镜观察该物质作用B16后的亚细胞结构变化。实验结果表明该物质可能为常温下性质较稳定的蛋白质类化合物，纯化到的单一组分WGF5对B16，Hela，MCF-7，Hep-3B肿瘤细胞均具有较强的抑制作用，其作用48 h的 IC₅₀（最低半抑制浓度）分别为2.767 μg/ml、2.204 μg/ml、3.758 μg/ml、3.073 μg/ml。MTT实验以及激光共聚焦显微镜下观察分析发现，蛋白WGF5能够明显改变细胞形态并最终导致肿瘤细胞死亡。从粘细菌Myxococcus macrosporus STXZ54分离到的活性物质具有广谱高效的抗肿瘤效果，有开发成抗肿瘤新药物的潜在价值。

关键词： 粘细菌; 抗肿瘤; 色谱分离; 激光共聚焦显微镜

Abstract:

Myxobacteria are known to be the third category of bacteria, which produce metabolites of potential anticancer effects. Here the lab-stored Myxococcus macrosporus STXZ54 was cultured, and components of its supernatants were attained by ammonium sulfate or acetone precipitation. Cytotoxic effects of the sediments were evaluated through an in vitro method. Further, the unique protein, WGF5, was successfully separated by high performance liquid chromatography from the myxobacteria. Its inhibitory effects on cancer cells were also analyzed by MTT assay. Simultaneously, confocal microscopy was employed to observe the alterations of B16 subcellular structure after treatment by WGF5. The results demonstrated that WGF5 was a very stable protein under room temperature and could efficiently inhibit the growth of B16, Hela, MCF-7 and Hep-3B cancer cells. The IC₅₀ of WGF5 after 48 h incubation with the cancer cells was 2.767 μg/ml, 2.204 μg/ml, 3.758 μg/ml and 3.073 μg/ml, respectively. Moreover, MTT assay and confocal microscopy observation found that WGF5 probably restrains the proliferation of the cancer cells through changing the cell morphology. Briefly, this study indicates that the WGF5 of Myxococcus macrosporus STXZ54 has an broad-spectrum anticancer effects and could be used for cancer therapy in the near future.

Key words: Myxobacteria Isolation Antitumor Confocal laser scanning microscope

收稿日期: 2014-05-29 出版日期: 2014-09-25

ZTFLH:

Q789

基金资助:

国家“863”计划（2011AA10A203）；国家自然科学基金（31070006）；湖南省生物发育工程及新产品研发协同创新中心（20134486）资助项目

通讯作者: 夏立秋 E-mail: xialq@hunnu.edu.cn

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引用本文:

文也, 唐少军, 肖蓉, 丁学知, 黄同龙, 雷良欢, 夏立秋. 粘细菌Myxococcus macrosporus STXZ54抗肿瘤活性物质的分离制备及其活性测定[J]. 中国生物工程杂志, 2014, 34(9): 63-71.

WEN Ye, TANG Shao-jun, XIAO Rong, HUANG Tong-long, LEI Liang-huan, DING Xue-zhi, XIA Li-qiu. Separation and Evaluation of Anticancer Components from Myxococcus Macrosporus STXZ54. China Biotechnology, 2014, 34(9): 63-71.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140910 或 https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I9/63

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