Effect of delta-elemene on Hela cell lines by apoptosis induction.
This study was designed to investigate the apoptosis-inducing activity of delta-elemene on Hela cells in vitro. MTT assay and Hoechst 33258/PI fluorescence microscopy were used for this investigation. Apoptosis was further confirmed and quantified by DNA fragmentation ELISA, Annexin V (AnV) binding of externalized phosphatidylserine and the mitochondrial probe JC-1 using flow cytometry. Generation of reactive oxygen species (ROS) was detected using CM-H2DCFDA. Western blots analysis was performed using antibodies against the pro-caspase-3, or PRAP (Poly (ADP-ribose) polymerase). The results showed that delta-elemene exhibited a marked antiproliferative effect on Hela cells in dose- and time-dependent manners, and had little inhibition to normal human liver cell line WRL-68. It was demonstrated that delta-elemene was capable of inducing DNA fragmentation in a dose- and time-dependent manner. AnV positivity and the disturbance of the polarized mitochondrial transmembrane potential (Deltapsim) suggested that delta-elemene induced apoptotic death of Hela cells. Western blot analysis demonstrated that delta-elemene activated the caspase-signaling pathway, leading to the proteolysis conversion of pro-caspase-3 to activate caspase-3, and the subsequent cleavage of the caspase substrate PARP. Further, it was noted that the apoptotic effect of delta-elemene could be attenuated by L-Glutathione (GSH) or z-DEVD-fmk. It suggested that the increase in ROS generation might be involved in the mechanism of delta-elemene induced cell apoptosis.
Wang
Xi-Sha
XS
Department of Pharmacology, Shenyang Pharmaceutical University, Liaoning Province, China, and Department of Paediatric, Prince of Wales Hospital, Chinese University of Hong Kong.
Yang
Wei
W
Tao
Shu-Juan
SJ
Li
Karen
K
Li
Ming
M
Dong
Jin-Hua
JH
Wang
Min-Wei
MW
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009842:Oligopeptides / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012717:Sesquiterpenes / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D013268:Stimulation, Chemical
D009842:Oligopeptides; D017382:Reactive Oxygen Species; D012717:Sesquiterpenes; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; C038905:elemene; D011065:Poly(ADP-ribose) Polymerases; D053148:Caspase 3; D005978:Glutathione