Investigation On Active Constituents Of Phellinus Linteus And The Molecular Mechanism For Its Anti-metastasis Effect

Phellinus linteus (PL) is a basidiomycete fungus that has been practiced in oriental countries for centuries to prevent ailments as diverse as gastroenteric dysfunction, diarrhea, haemorrhage and cancers. The fall in supply and the increase in demand have stimulated interest in the search for substitutes of the natural products of the fungus. Although it is wildly believed in Asia that the interplay of different constituents of PL is responsible for its health-promoting effects, this notion, among many other theories of traditional Asian medicine, is mainly established on empirical clinical observations and lacks credibility proved by the contemporary medical science. These questions hamper the development of PL by modern pharmaceutical programs. Aimed on these questions, we started the studies as follows:1. This paper describes a submerged culture approach that provides abundant natural flavonoids from a medicinal mushroom Phellinus linteus, and the optimal conditions of flavonoids productivity was determined in culture broth containing different nutrition sources, especially soybean cake. An optimal medium composition (3%glucose,2%lactose,1%beef extract,0.3%soybean cake,0.1%KH2PO4, and0.05%MgSCO4) for high flavonoids productivity. The addition of soybean cake could improve flavonoids productivity, which is4times higher than basic media. Furthermore, Antioxidant activity of flavonoids from soybean cake culture was higher compared to that of flavonoids from the culture without soybean cake, but its antitumor activity was lower.2. The chemical constituents of submerged Phellinus linteus were investigated.13compounds, including2new straight chain type sesquiterpenes, were isolated from culture broth of Phellinus linteus by varied chromatographic methods, and their structures were established on the basis of extensive spectroscopic analysis (including1D and2D NMR, MS, HR-MS, IR). The volatile components acetoacetate extractant of submerged Phellinus linteus were analyzed by gas chromatography-mass spectrometry.13componds were identified, which included ergosterol, polyunsaturated fatty acid (PUFA) and cyclic dipeptides, which were frist identified from Phellinus linteus.3. The anticancer potential of Hispolon was further explored by examiningits efficacy against phorbol ester (TPA)-stimulated Epithelial Mesenchymal Transition (EMT) in breast cancer cell. Hispolon was demonstrated to significantly inhibit TPA stimulated migration of MCF-7cells in an in vitro transwell assay. The inhibition oftumor cell migration by Hispolon was found to occur at the transcriptional level through the inhibition of E-cadherin. It was thus concluded that by targeting TPA stimulated significant increase in the levels of intracellular ROS, Hispolon inhibited MCF-7cell migration through decreasing the phosphorylation levels of ERK, which in turn inhibit the expression of transcription inhibition factor Slug, eventually induced reexpression of E-cadherin. These findings presented in this study had expanded on the current understanding of the anticancer potential of Hispolon from Phellinus linteus.Taken together, these findings will contribute to the search for substitutes of the natural products of the medicinal mushroom, and exploring the molecular mechanism of active components from the medicinal mushroom. Moreover, this study could be useful for the drug development from traditional Chinese medicinal mushrooms.