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Two steroidal saponins from Camassia cusickii induce L1210 cell death through the apoptotic mechanism

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Two steroidal saponins, tigogenin hexasaccharide-1 (TGHS-1, (25R)-5α-spirostan-3-yl 4-O-[2-O-[3-O- (α-L-rhamnopyranosyl)--D-glucopyranosyl]-3-O-[4-O-(α-L-rhamnopyranosyl)--D-glucopyranosyl]--D-glucopyranosyl]- -D-galactopyranoside) and tigogenin hexasaccharide-2 (TGHS-2, (25R)-5α-spirostan-3-yl 4-O-[2-O-[3-O- (-D-glucopyranosyl)--D-glucopyranosyl]-3-O-[4-O-(α-L-rhamnopyranosyl)--D-glucopyranosyl]--D-glucopyranosyl]- -D-galactopyranoside), were isolated from the fresh bulbs of Camassia cusickii. In murine leukemic L1210 cells, both compounds showed cytotoxicity with an EC50 value of 0.06 µM. The morphological observation revealed that TGHS-1 and TGHS-2 induced shrinkage in cell soma and chromatin condensation, suggesting apoptotic cell death. The cell death was confirmed to be apoptosis by Annexin V binding to phosphatidylserine in the cell membrane and excluding propidium iodide. A typical apoptotic DNA ladder and the cleavage of caspase-3 were observed after treatment with TGHS-1 and TGHS-2. In the presence of both the compounds, cells with sub-G1 DNA content were detected by flow cytometric analysis, indicating that TGHS-1 and TGHS-2 (each EC50 value of 0.1 µM) are the most powerful apoptotic saponins known. These results suggest that TGHS-1 and TGHS-2 induce apoptotic cell death through caspase-3 activation.Key words: steroidal saponin, tigogenin hexasaccharide, apoptosis, DNA fragmentation, murine leukemic L1210 cells.

Les deux saponines stéroïdiennes, tigogénine hexasaccharide-1 (TGHS-1, (25R)-5α-spirostan-3-yl 4-O- [2-O-[3-O-(α-L-rhamnopyranosyl)--D-glucopyranosyl]-3-O-[4-O-(α-L-rhamnopyranosyl)--D-glucopyranosyl]- -D-galactopyranoside) et tigogénine hexasaccharide-2 (TGHS-2, (25R)-5α-spirostan-3-yl 4-O-[2-O-[3-O- (-D-glucopyranosyl)--D-glucopyranosyl]-3-O-[4-O-(α-L-rhamnopyranosyl)--D-glucopyranosyl]--D-glucopyranosyl]- -D-galactopyranoside), ont été isolées de bulbes frais du Camassia cusickii. Dans des cellules leucémiques L1210 murines, les deux composés ont montré une cytotoxicité avec une CE50 de 0,06 µM. L'observation morphologique a révélé que TGHS-1 et TGHS-2 ont entraîné une réduction du corps cellulaire avec condensation de la chromatine, suggérant une mort cellulaire programmée. La mort cellulaire par apoptose a été établie par la fixation d'annexine V à la phosphatidylsérine dans la membrane cellulaire et par l'exclusion à l'iodure de propidium. Une échelle d'ADN apoptotique typique et le clivage de la caspase-3 ont été observés après le traitement avec TGHS-1 et TGHS-2. En présence des deux composés, des cellules ayant une teneur en ADN sub-G1 ont été détectées à l'analyse par cytométrie en flux, ce qui indique que TGHS-1 et TGHS-2 (CE50 de 0,1 µM) sont les deux plus puissantes saponines apoptotiques connues. Ces résultats permettent de présumer q

Keywords: DNA fragmentation; apoptose; apoptosis; cellules leucémiques L1210 murines; fragmentation de l'ADN; murine leukemic L1210 cells; saponine stéroïdienne; steroidal saponin; tigogenin hexasaccharide; tigogénine hexasaccharide

Document Type: Research Article

Publication date: November 1, 2001

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