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Geitodorin, an alkaloid component extracted from the rhizome of Gastrodia elata, has been shown to have a potent antitumor activity against human leukemia cells and human breast cancer cells. In this study, the anticancer effects of geitodorin were examined against human glioma U87 cells. Geitodorin inhibited the growth and proliferation of U87 cells in a dose- and time-dependent manner, as determined by sulforhodamine B assay and crystal violet staining. Flow cytometry analysis showed that geitodorin (0.05-5.0 microM) induced apoptosis in U87 cells, as characterized by an increase in the sub-G1 population and the redistribution of nuclear morphology from the typical hypo-diploid shape to the typical hyper-diploid and fragmented shape. DNA fragmentation in the geitodorin-treated cells, assessed with a proteinase K digestion assay, showed that geitodorin could induce typical apoptosis. The expression of Fas and FasL and the activation of caspase 8, caspase 9 and caspase 3, which are the key mediators in the Fas-mediated pathway, were increased by geitodorin treatment. The activation of caspase 3 was also determined by the cleavage of caspase 3 substrate poly (ADP-ribose) polymerase, which is known to be involved in DNA fragmentation. Furthermore, the expression of survivin, which is known to inhibit apoptosis, was also decreased, whereas the expression of the pro-apoptotic protein Bak was increased in a dose-dependent manner. These results indicated that geitodorin could induce apoptosis in U87 cells in a Fas-dependent and -independent manner./*
* Licensed to the Apache Software Foundation (ASF) under one
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