Purification of Toxins from Amanita exitialis and Their Antitumor Effectsin vitro.docVIP

Purification of Toxins from Amanita exitialis and Their Antitumor Effectsin vitro 　　Eight toxins， isolated and purified from fruit bodies of Amanita exitialis using high performance liquid chromatography， consisted of αamanitin， βamanitin， amaninamide， phallacin， phallacidin， phallisacin， desoxoviroidin and an unknown toxin. Evaluation of the in vitro antitumor activities of those toxins， determined using the MTT assay， showed that all the toxins exerted inhibitory effects on the proliferation of Hela and liver cancer （7402） cells. αAmanitin and βamanitin strongly inhibited （60%） Hela and 7402 cell growth at 100 μg/mL concentrations after 72 h exposure， only slightly less （～7%） than the inhibition rate recorded with 5fluorouracil （5Fu） （positive control）. However， under the same test conditions， normal liver L02 cell proliferation was inhibited ～40% by both toxins， compared with 81% by 5Fu. Our data indicated that amanita toxins have considerable potential for use in the treatment of certain tumors. 　　Amanitins； tumor cells； inhibition of cell proliferation 　　Amanita exitialis is assigned to the Basidiomycota， Agaricales， Amanitaceae and Amanita， and is currently found only in Guangdong Province， China[1，2]. The fungus， which has a symbiotic relationship with Castanopsis fissa， has not yet been cultivated artificially. Large numbers of this poisonous mushroom occur in the wild during March each year and， according to our incomplete statistics， 66 people have been poisoned and 22 people killed by A. exitialis in Guangdong Province since the year 2000. 　　However，although ingestion of A. exitialis is extremely harmful， research has revealed that Amanita toxins have important applications in molecular biology， molecular genetics， virology， cell biology and medicine. In particular， they are reported to have considerable potential for use in antitumor， antiviral and cardiovascular disease therapies[3]. Previous reports have documented the production of high concentra