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目的考察菌株Streptomyces capillispiralis 1070的代谢产物抑制肿瘤细胞A549增殖的作用机制。方法采用MTT法考察菌株Streptomyces capillispiralis 1070的活性代谢产物对A549细胞增殖的抑制作用,采用吖啶橙活细胞染色荧光显微镜观察法、罗丹明123活细胞染色流式细胞检测技术及扫描电镜技术、透射电镜技术观察A549细胞增殖受到抑制后的各种变化。采用全波长扫描与Molish反应对活性物质的成分做初步判断。结果该活性物质可使A549细胞存活率降低至25.79%,可使A549细胞线粒体膜电位明显下降,能使A549细胞胞浆内出现酸性自噬泡,但细胞膜结构完整、无细胞内容物外漏。结论推测该活性物质是通过诱导细胞自噬而发挥其抗肿瘤作用。菌株Streptomyces capillispiralis 1070的活性代谢产物为多糖类物质。
Objective To investigate the mechanism of the inhibition of the proliferation of tumor cell A549 by the metabolites of strain Streptomyces capillispiralis 1070. Methods The inhibitory effect of active metabolites of strain Streptomyces capillispiralis 1070 on the proliferation of A549 cells was investigated by MTT method. The fluorescence cell microscopy of acridine orange live cells staining, flow cytometry using rhodamine 123 live cell staining, scanning electron microscopy, and transmission were used. Electron microscopy technique was used to observe various changes after A549 cell proliferation was inhibited. Full-wavelength scanning and Molish reactions were used to make preliminary judgments on the composition of the active material. RESULTS: The active substance could reduce the survival rate of A549 cells to 25.79%. The mitochondrial membrane potential of A549 cells was significantly decreased, and acidic autophagic vacuoles appeared in the cytoplasm of A549 cells. However, the cell membrane structure was intact and there was no leakage of cell contents. Conclusion It is speculated that the active substance exerts its anti-tumor effect by inducing autophagy. The active metabolite of strain Streptomyces capillispiralis 1070 is a polysaccharide material.