Apoptosis and cytolysis induced by giganteosides and hederacolchisides in HL-60 cells.
The viability, cytolysis and apoptosis-mediated cellular death induced by giganteosides D and E (Gig-D and Gig-E) and hederacolchisides A and A1 (Hcol-A and Hcol-A1) were analysed in HL-60 cells.


METHODS
The end-point metabolic (WST1) and lactate dehydrogenase (LDH) assays were used. Cell cycle analysis and apoptosis were measured by flow cytometry, DNA laddering and caspase-3 analyses.


RESULTS
the HL-60 cell line was more sensitive to Hcol-A1 and Gig-D (IC50 3-5 microM) than to Gig-E and Hcol-A (IC50 8-13 microM; WST1 assay). This was related to LDH release. The induction of apoptosis could be detected without caspase-3 activation after 24 h of treatment. DNA fragmentation could be detected only with Gig-D. With Hcol-A1 and Gig-D, an accumulation of cells in the S-phase and an increase of cells in sub-G1 peak were observed. By the annexinV-fluorescein isothiocyanate (FITC)/7-amino-actinomycin D (AAD) assay, the majority of cells were in late apoptosis with Gig-D, and in necrosis with Hcol-A1.


CONCLUSIONS
Hcol-A1 is more cytotoxic than Gig-D, followed by Gig-E and finally Hcol-A. This is related to a membrane permeabilization effect, leading to cytolysis.
Gerkens
Pascal C
PC
Laboratories of Mass Spectrometry and Histology - Cytology (GIGA-CRCE), Institute of Chemistry (B6c), University of Liège, Sart - Tilman, B-4000 Liège 1, Belgium.
Dobson
Rowan
R
Tabatadze
Nino
N
Mshviladzade
Vakhtang
V
Elias
Riad
R
Peulen
Olivier J
OJ
Jolois
Olivier M
OM
De Pauw-Gillet
Marie-Claire A
MC
D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D048708:Cell Growth Processes / Q000187:drug effects; D002463:Cell Membrane Permeability; D049917:Dipsacaceae / Q000737:chemistry; D018922:HL-60 Cells; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology; D012503:Saponins / Q000494:pharmacology*
D010936:Plant Extracts; D012503:Saponins; C467827:hederacolchisid A1; C412406:hederacolchiside A; D007770:L-Lactate Dehydrogenase