Analysis of Antrodia cinnamomea biological activity and the terpinoid biosynthesis related genes
Date Issued
2005
Date
2005
Author(s)
Tong, Kai-Hong
DOI
zh-TW
Abstract
Antrodia cinnamomea is a fungus endogenous to Taiwan and thought to have anticancer activities. In current study we attempt to analyze and elucidate the genes that possibly involved with terpenoids biosynthesis. Six putative genes：ergosterol biosynthesis protein，Putative mevalonate kinase，Diphosphomevalonate decarboxylase，Farnesyl cysteine carboxyl methyltransferase,squalene epoxidase and Lanosterol 14 α-demethylase , were identified from the A. cinnamomea cDNA library after blastX with nr NCBI database, and primers were designed to amplify these specific genes using gDNA of A. cinnamomea as template by polymerase chain reaction. The obtained gene products were further sequenced, and their identies verified, labeled with DIG and used as probe to screen the constructed A. cinnamomea Fosmid library. The results from colony hybridization revealed these genes were not allocated in a cluster. In order to investigate further on the possibility of terpenoids in anticancer activity, we used flucanazole, butenafin, and compactin, which are thought to block terpenoids biosynthesis pathway at specific steps. The effect of these inhibitors on A. cinnamomea were analyzed using High Performance Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC) in combination with the bioactivity assay using liver cancer cell Hep G2 and lung cancer cell A549. The results showed both the biosynthesis of terpenoids and the anticancer activity of A. cinnamomea were not significantly affected by compactin; with respect to butenafin, through the terpenoids varied greatly either qualitatively or quantitatively, the anticancer activity was not changed in a regular pattern; while flucanazole markedly reduced the biosynthesis of terpenoids as well as the anticancer activity. Consequently, it is suggested that the downstream terpenoids products mediated by lanosterol 14α-demethylase was mainly ascribed to the anticancer activity. Currently,we are conducting lanosterol 14 α-demethylase gene silencing by using siRNA to prove the actual role played by these gene products in suppression of the liver cancer cell Hep G2.
Subjects
牛樟芝
三萜
類
Antrodia cinnamomea
tritenpene
SDGs
Type
other
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