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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IC_(50)(最低半抑制浓度)分别为2.767μg/ml、2.204μg/ml、3.758μg/ml、3.073μg/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。
Myxobacteria as the third largest secondary metabolites producing bacteria, is an important resource for mining new anti-tumor drugs. After the myxococcus macrosporus STXZ54 was preserved in our laboratory, the fermentation broth was subjected to ammonium sulfate precipitation and acetone precipitation. The properties of the antitumor active substances in the fermentation broth of the strain were analyzed. High performance liquid chromatography The antitumor active substances in the fermentation broth were separated, and the cytotoxicity of the substance was measured. The changes of the subcellular structures after the action of B16 were observed by laser confocal microscopy. Experimental results show that the substance may be a stable protein compound at room temperature. The purified single component WGF5 has a strong inhibitory effect on B16, Hela, MCF-7 and Hep-3B tumor cells, and its effect for 48 h IC 50 (lowest half inhibitory concentration) were 2.767μg / ml, 2.204μg / ml, 3.758μg / ml and 3.073μg / ml, respectively. MTT experiments and confocal laser scanning microscopy analysis found that the protein WGF5 can significantly change the cell morphology and eventually lead to tumor cell death. The active substances isolated from the myxococcus macrosporus STXZ54 have a broad spectrum of highly effective antitumor effects and potential value as new antitumor drugs.