Interaction of Ganoderma triterpenes with doxorubicin and proteomic characterization of the possible molecular targets of Ganoderma triterpenes.
Triterpenes are the main components with cytotoxicity in Ganoderma lucidum, which is used popularly as a complementary treatment for cancer therapy in traditional Chinese medicine. To investigate the possible interaction between chemotherapeutic agents and triterpenes extracted from G. lucidum, the cytotoxicity of doxorubicin (DOX) combined with Ganoderma triterpenes (GTS) or lucidenic acid N (LCN), a purified compound, was examined in HeLa cells. The combinations targeting DOX with GTS or LCN resulted in a synergistic interaction in HeLa cells. Moreover, to identify the molecular targets of GTS, two-dimensional gel electrophoresis-based comparative proteomics was carried out and proteins with altered expression levels after GTS treatment in HeLa cells were identified by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. The results of our proteomic study indicated that the GTS treatment caused regulated expression of 14 proteins, which play important roles in cell proliferation, the cell cycle, apoptosis, and oxidative stress. Flow cytometric analysis confirmed that GTS could induce weak G(0)-G(1) phase arrest and combined use of GTS with DOX could induce apoptosis in cells. Furthermore, GTS enhanced the reactive oxygen species (ROS)-producing effect of DOX, and a ROS scavenger could affect the synergism between GTS and DOX. In cells with high Ku80 protein expression, the synergism between GTS and DOX was also partly affected. Importantly, in cells with high Ku80 expression that were treated with a ROS scavenger, the synergism between GTS and DOX totally disappeared. These results suggest that the synergism between GTS and DOX might be based on GTS-induced sensitization of cells to chemotherapeutics through enhanced oxidative stress, DNA damage, and apoptosis.
Yue
Qing-Xi
QX
Shanghai Research Center for Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Xie
Fu-Bo
FB
Guan
Shu-Hong
SH
Ma
Chao
C
Yang
Min
M
Jiang
Bao-Hong
BH
Liu
Xuan
X
Guo
De-An
DA
D034961:Antigens, Nuclear / Q000032:analysis; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000032:analysis; D004317:Doxorubicin / Q000494:pharmacology*; D004357:Drug Synergism; D032902:Ganoderma / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D000072200:Ku Autoantigen; D009363:Neoplasm Proteins / Q000032:analysis*; D054648:Protein Phosphatase 2 / Q000502:physiology; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism; D014315:Triterpenes / Q000494:pharmacology*
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009363:Neoplasm Proteins; D017382:Reactive Oxygen Species; D014315:Triterpenes; C437028:lucidenic acid N; D004317:Doxorubicin; C517189:PPP2CA protein, human; D054648:Protein Phosphatase 2; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen