真菌PHF-9化学成分及其抗肿瘤活性研究

摘要

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Exopisiod 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89.5 μmol·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 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 spectro-scopic 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 Exopi-siod l,were purified from the acetic ether extract of fungus strain PHF-9. Exopisiod 1 exhibited moderate cytotoxici-ties 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 |xmol-L , 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.