| Objective: We had separated a new source of deep-sea Aspergillus sp previous.Here, we aim to extract, purify the secondary metabolites and identify their structure.And then study the cytotoxic activity of the isolated compounds on hepatic carcinomacells and preliminary study its anticancer mechanism.Methods: After small-scale fermentation, secondary metabolites were extractedwith ethyl acetate. Chromatography was used to separate and purify the extraction.Identify the extractive with MS、NMR. Cytotoxic of the new identified compoundson hepatic carcinoma cells was measured by CCK-8assay. Protein expression wasdetermined using two-dimensional gel electrophoresis to preliminary study themechanism of the cytotoxic activity.Result： Four novel compounds were obtained from the secondary metabolite.They are sterigmatocystine,anhydroorcinol,1’-methoxy-averantin, and averufine.Anhydroorcinol posseses the strongest cytotoxic activity to HCC, but low cytotoxicactivity to the normal liver cells. Compare the result of2DE in cancer and normalcells,12proteins spots were changed significantly. Among of the12spots,8spotsare common owned of two groups where2spots are up-regulated and6spots aredown-regulated in Bel7402group. Use MALDI-TOF/TOF-MS to identify the8spots,obtain6cerdible results(p <0.05). Six protein spots are KRT8, Keratin, type Icytoskeletal19, Keratin, type I cytoskeletal10、Alpha-enolase isoform1、Glucose-6-phoaphate1-dehydrogenase isoform b, there is an unknown protein(protein for IMAGE:3534054).Conclusion：1, Four new compounds were sepreted and purified from thesecondary metabolites of Aspergillus, of which the anhydroorcinol inhibitedhepatomacell cytotoxic activity, cytotoxic activity was lower for normal liver cells.2, After compound anhydroorcinol act on the HCC cells,, there are12protein spots are changed significantly, including8spots are common owned in normal cells andHCC cells where2protein spots were up-regulated,6spots down-regulated. |
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