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Juglone-induced apoptosis in human gastric cancer SGC-7901 cells via the mitochondrial pathway.

Abstract
This study was designed to investigate the effect of juglone on the apoptosis of human gastric cancer SGC-7901 cells. The cytotoxic activity of juglone on SGC-7901 cells was tested by the sulforhodamine B (SRB) assay. The morphological changes in the cells were observed by transmission electron microscopy (TEM). The apoptotic rate, the level of reactive oxygen species (ROS), mitochondrial transmembrane potential and the expression of cytochrome c protein were detected by flow cytometry (FCM). The expression of Bcl-2 and Bax proteins were examined by Western blot. Caspase 3 activity was determined with a microplate reader. Our results were as follows: the GI(50) values for SGC-7901 cells were 36.51 ± 1.05 μmol/L (24h) and 25.37 ± 1.19 μmol/L (48 h). After 24h of exposure to juglone (5, 10, 15 and 20 μmol/L), the cells presented the typical morphological changes of apoptosis, and the rate of apoptosis was found to increase in a dose-dependent manner. After cells were treated with juglone at the same dose for 24h, the level of ROS was significantly higher, the expression of Bcl-2 was significantly down-regulated and the expression of Bax was significantly up-regulated compared to the control. The mitochondrial transmembrane potential was significantly lower, and the expression of the cytochrome c protein was significantly higher relative to the control. Caspase 3 was activated in a concentration-dependent manner. In conclusion, juglone can induce apoptosis in SGC-7901 cells through a mitochondrial pathway that seems to be mediated by the generation of ROS and a reduction in the Bcl-2/Bax ratio.
AuthorsYu-Bin Ji, Zhong-Yuan Qu, Xiang Zou
JournalExperimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie (Exp Toxicol Pathol) Vol. 63 Issue 1-2 Pg. 69-78 (Jan 2011) ISSN: 1618-1433 [Electronic] Germany
PMID19815401 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCrown Copyright © 2009. Published by Elsevier GmbH. All rights reserved.
Chemical References
  • Antineoplastic Agents
  • BAX protein, human
  • Naphthoquinones
  • Proto-Oncogene Proteins c-bcl-2
  • Reactive Oxygen Species
  • bcl-2-Associated X Protein
  • Cytochromes c
  • Caspase 3
  • juglone
Topics
  • Antineoplastic Agents (pharmacology)
  • Apoptosis (drug effects)
  • Blotting, Western
  • Caspase 3 (metabolism)
  • Cell Line, Tumor
  • Cytochromes c (biosynthesis)
  • Humans
  • Membrane Potential, Mitochondrial (drug effects)
  • Microscopy, Electron, Transmission
  • Mitochondria (drug effects, ultrastructure)
  • Naphthoquinones (pharmacology)
  • Proto-Oncogene Proteins c-bcl-2 (biosynthesis)
  • Reactive Oxygen Species (metabolism)
  • Stomach Neoplasms (metabolism, pathology, ultrastructure)
  • bcl-2-Associated X Protein (biosynthesis)

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