真菌PHF-9化学成分及其抗肿瘤活性研究

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。

Chemical metabolites from fungus PHF-9 and their anti-tumor activity

Objective:To isolate and identify the cytotoxic metabolites from the soil-derived fungus strain PHF-9 and elucidate the possible mechanism of their anti-tumor action.Methods:The acetic ether extract was separated by combined chromatography methods.The isolated compounds were identified by analysis of the spectroscopic data.MTT assay was used to evaluate the inhibition of certain cancer cell lines by the compounds.The morphological changes were observed by Hoechst 33342 staining.Annexin V/PI assay was performed to determine their apoptotic effects.Results:Four compounds including a new naturally occurring indole alkaloid,designated Exopisiod 1,were purified from the acetic ether extract of fungus strain PHF-9.Exopisiod 1 exhibited moderate cytotoxicities against A-549,Hela,PANC-28 and BEL-7402 cell lines,with the IC50 values of 76.3,77.2,107.5 and 89.5 μmol·L-1,respectively.Exposure of A549 cancer cells to Exopisiod 1 resulted in some morphological changes associated with apoptosis.Additionally,the proportion of early apoptotic cells increased in a dose-dependent manner.Conclusion:Soil-derived fungus strain Exophiala pisciphila can produce indole alkaloid,which inhibits the growth of cancer cells through induction of apoptosis.