Anti-tumor activity of Gastrodia elata Blume is closely associated with a GTP-Ras-dependent pathway

Abstract

Abstract. Gastrodia elata Blume (GEB) is an important medicinal plant in Korea. In order to confirm the anti-tumor activities of GEB extracts, we carried out various in vitro antitumor assays, including a wound assay and an invasion assay using an ethyl ether extract of GEB. The results showed that the GEB extract exhibits potent anti-tumor activity in vitro in a dose-dependent manner. The expression of CD44, cdc42, Timp-2 or RhoA mRNA did not change by GEB treatment, compared to that of the control. GTP-Ras, an active form of a G-coupled protein family, however, is associated with the anti-tumor activity of GEB extracts. We examined various molecular markers related to metastasis by reverse transcriptase-polymerase chain reaction with the extract of GEB-treated B16 cells. There was an increase in GTP-Ras expression by the Gastrodia elata Blume extract. Together, these results suggest that the Gastrodia elata Blume extract could have potential in alleviating tumorigenesis, by a GTPRas-dependent pathway; although the precise molecular mechanisms are still being examined. IntroductionGastrodia elata Blume (GEB) is a traditional herb that has been used in East Asian for centuries. It has been used as an anticonvulsant, analgesic and sedative to combat vertigo, hypertension, general paralysis and tetanus. Vanillyl alcohol and gastrodin, derived from GEB are known to have anticonvulsive actions (1). Recently, it was reported that compounds found in GEB inhibited glutamate-induced apoptosis in neuronal cells (2). In addition, after pentylenetetrazole-induced seizure activity, the ether fraction of GEB has been shown to attenuate a decrease in γ-aminobutyric acid (GABA) and an increase in glutamate content, as well as having anticonvulsant effects (3).GEB is also used as sub-material for food or food-related industry. In 2001, the Korea Food and Drug Administration approved that GEB extracts as food ingredients; however, an in vivo toxicological study is still needed to determine its safety in food. We have already reported that GEB powersupplemented (0.5-1.0%) dough had a membrane-like structure that was more developed than that of the control, resulting in increased bread volumes (4). These results suggest that bread baked with 0.5-1.0% GEB exhibited an increase in loaf volume due to the more complete development of a gluten matrix.In our study, we determined that GEB protected cell damage by ß-amyloid in neuroblastoma cells (5). The ethyl ether fraction of GEB has potent activity toward ß-amyloidinduced cell damage via reducing caspase-3 activity (6,7; data not shown). In the course of a mechanistic study of the methanolic extract of GEB, we found that it has potent antitumor activity in vitro. The major finding of this report is that GEB exhibits anti-metastatic activities which were confirmed by wound and invasion assay in B16 melanoma cells. We further demonstrated that GEB increased anti-tumor activity via a GTP-Ras-dependent pathway. Materials and methodsCell culture. A murine melanoma cell line B16-F1 (B16; Ca...

Protective effects of Gastrodia elata Blume on MPP+-induced cytotoxicity in human dopaminergic SH-SY5Y cells

“…It is also used for the treatment of inflammatory diseases, headache, epilepsy, and other neurological disorders (Tang and Eisenbran, 1992;Kim et al, 2001). Recently, GE was reported as an important medical food and some of the active constituents possess anti-atherosclerotic properties Lee et al, 2009), anti-angiogenic activity (Ahn et al, 2007), anti-tumor activity (Heo et al, 2007). Its beneficial effects in central nervous system were also documented (Liu and Mori, 1992;Hsieh et al, 1997;Kim et al, 2003;Sun et al, 2004;Jung et al, 2006Jung et al, , 2007Huang et al, 2007).…”

A fraction of methylene chloride from Geum japonicum Thunberg inhibits tumor metastatic and angiogenic potential

“…Attachment and spreading assays. Attachment and spreading assays were carried out as previously described (9). Briefly, the B16 cells spread to the bottom of each well of the 6-well plates (Greiner, Frickenhausen, Germany) at a concentration of the GjT extract (1x10 4 cells/well) for 2 h (attach assay) or 24 h (spreading assay) in a 5% CO 2 incubator at 37˚C.…”

“…The oligo nucleotides were used in PCR amplification with specific primers for mice CD44 (forward, 5'-TCG ATT TGA ATG TAA CCT GCC-3'; reverse, 5'-TGG TGT GTT CTA TAC TCG CCC-3'), TIMP-2 (forward, 5'-GCA TCA CCC AGA AGA AGA GC -3'; reverse, 5'-GGG TCC TCG ATG TCA AGA AA-3') and GAPDH (forward, 5'-ATG TTC CAG TAT GAC TCC AC -3'; reverse, 5'-GCC AAA GTT GTC ATG GAT GA-3') as an internal control. The PCR was carried out for the products of CD44, TIMP-2 and glyceraldehydes-3-phosphate dehydrogenase (GAPDH), with denaturing at 94˚C for 30 sec, annealing at 58˚C for 30 sec and extension at 72˚C for 30 sec (9). The linear quantitation of the RT-PCR products was determined by a Gel-doc System (Bio-Rad, Universal-II, San Jose, CA, USA).…”

Reduction of matrix metalloproteinase-9 expression by culture filtrate of Paecilomyces farinosus J3

“…transwell plates (pore size 8 µm; costar, ny, uSa) were loaded with 100 µl matrigel (Bd Biosciences), allowed to solidify at 37˚C for 2 h and then coated with 10 µl of fibronectin (200 µg/ml). The plates were loaded with B16 cells suspended in 10% FBS (1x10 5 cells/well), the samples were exposed to J3 (0-100 mg/ml) and the plates were incubated at 37˚C in 5% CO 2 for 24 h. the migrated cells were fixed with methanol, stained with hematoxylin and counted under a microscope (15).…”

“…this cell line is a good model for the assessment of cell morphology, motility and proliferation (13)(14)(15). B16 cells migrated into the damaged wound, suggesting that migration is associated with various events in the cytoskeletal structure, such as actin polymerization and changes in the cell motility-related framework (30).…”