Study Of Anticancerous Triterpenes From Mycelia Of Twenty-six Ganoderma Strains

This study was on the triterpene extracts from liquid fermentation mycelia of 26 strains falling into 5 different species of Ganoderma genera (G. lucidum, G. tsugae, G. oerstedii, G. subamboinense, G. resinaceum). First, the optimal conditions of vanillin-perchloric acid colorimetric reaction were studied for quantitative detection of total triterpenoid derived from Ganoderma. They were: to evaporate sample solvent (methanol) at 70℃, then the sample residue was reacted with 0.1 ml 5% (w/v) vanillin in glacial acetic acid and 1.0 ml perchloric acid at 60℃in water bath for 15 minutes. Afterwards, 3.9ml glacial acetic acid was added to terminate the reaction and the optical density was measured at 547 nm. Next, MTT assay was used to study the in vitro anti-proliferation activities of the crude triterpene (GLF) extracted with ethanol and its acidic fraction (GLT) and none acidic fraction (GLX). The results showed that GLF significantly inhibited proliferation of human acute promyelocytic leukemia cells HL-60 and human breast cancer cells MCF-7. The percentages of inhibition were 65.4+3.6% (HL-60, 150μg/ml) and 97.8+7.4% (MCF-7, 400μg/ml), respectively, after 72 hours treatment. Such inhibitory effect of GLT was much weaker (16.4+4.8% and 33.3+10.1% respectively for HL-60 and MCF-7), compared to GLF. In contrast, GLX showed a stronger potency than GLF, while its percentages of inhibition were 82.5+8.9% and 98.0+7.9% (HL-60 and MCF-7) respectively. The morphological observation and Annexin V flow cytometric assay revealed that both GLF and GLX at 100μg/ml could induce the apoptosis in HL-60 cells while GLT didn't exert such activity. Therefore, the crude triterpene extracts, instead of its purified fractions, were used in the following screening tests. By employing the above mentioned vanillin colorimetric method and MTT assay, triterpene extracts from liquid fermentation mycelia of the 26 different strains were studied on their total triterpene contents and anti-proliferation activities against HL-60 and MCF-7 in vitro. The results showed that all the strains mycelia produced triterpenes, while their total triterpene contents were from 8.20±0.44% to15.85±0.07% in the extracts. Among these, the triterpene content in T3 strain sample was the highest (15.85±0.07%). These triterpene extracts all exerted anti-proliferation activities against HL-60 cells within a concentration range from 50μg/ml to 250μg/ml and against MCF-7...