论文部分内容阅读
目的:分离鉴定土壤来源的真菌菌株PHF-9中的抗肿瘤活性成分并研究其抗肿瘤机制。方法:采用多种色谱方法对乙酸乙酯提取物中的化学成分进行分离,并通过波谱数据鉴定化合物结构;通过MTT法测定细胞增殖抑制率,Hoechst 33342染色法观察细胞形态变化,Annexin V/PI双染法分析细胞凋亡。结果:从菌株PHF-9的发酵产物中分离得到4个化合物,包括一个吲哚生物碱类新天然产物,命名为Exopi-siod 1。MTT检测显示Exopisiod 1对A-549,Hela,PANC-28和BEL-7402肿瘤细胞株的IC50分别为76.3,77.2,107.5和89.5μmol·L-1。Exopisiod 1作用后的A549细胞在荧光显微镜下可见凋亡相关的形态学变化;Annexin V/PI双染法显示细胞凋亡百分率呈剂量依赖关系。结论:土壤来源的外瓶霉属真菌可以代谢具有抗肿瘤活性的吲哚生物碱,初步推断其通过诱导细胞凋亡抑制肿瘤细胞生长。
OBJECTIVE: To isolate and identify the antitumor components of fungal strain PHF-9 from soil and study its antitumor mechanism. Methods: The chemical constituents of ethyl acetate extract were separated by a variety of chromatographic methods. The structures of the compounds were identified by spectral data. The cell proliferation inhibition rate was determined by MTT assay. The morphological changes of cells were observed by Hoechst 33342 staining. Annexin V / PI Double staining analysis of apoptosis. Results: Four compounds were isolated from the fermentation product of strain PHF-9, including a new natural product of indole alkaloid named Exopi-siod 1. MTT assay showed that the IC50 of Exopisiod 1 in A-549, Hela, PANC-28 and BEL-7402 tumor cell lines were 76.3, 77.2, 107.5 and 89.5 μmol·L -1, respectively. A549 cells treated with Exopisiod 1 showed apoptosis-related morphological changes under the fluorescence microscope; Annexin V / PI double staining showed that the percentage of apoptosis was dose-dependent. CONCLUSION: The soil-derived Ectomycorrhizal fungi can metabolize indole alkaloids with antitumor activity, and presumably inhibit the growth of tumor cells by inducing apoptosis.