Effect of arsenic trioxide on uveal melanoma cell proliferation in vitro.
OBJECTIVE
The aim of this study was to investigate the effect of arsenic trioxide (ATO) on the growth of cultured uveal melanoma cells (OCM-1 cell line).


METHODS
Cultured OCM-1 cells were treated with 0.75-24 microM of ATO for 4-96 h. Cell viability was evaluated with the methylthiazoletetrazolium (MTT) assay as proliferation test. Apoptotic and necrotic cells were quantified using flow cytometry following Annexin-V/PI double stain. The cell morphology alteration was examined by light and electron transmission microscopy. To investigate the underlying mechanism of ATO-induced apoptosis and necrosis, glutathione peroxidase activity was measured, and mitochondrial membrane potentials were quantified using confocal microscopy.


RESULTS
In the MTT assays, OCM-1 cell growth was inhibited at ATO concentrations of 1.5-24 microM. The 50% inhibitory concentration of ATO was 16.8 microM. At ATO concentrations of 12 and 24 microM, apoptosis and necrosis were induced after 24 h of incubation as shown by light and transmission electron microscopy. At ATO concentrations of 12 and 24 microM, the glutathione peroxidase activity was significantly (p < 0.05) reduced after 24 h of incubation, and the mitochondrial membrane potentials were significantly (p < 0.01) decreased after 4, 12 and 24 h of incubation.


CONCLUSIONS
ATO inhibited ocular melanoma cell line growth at concentrations of 1.5-24 microM of ATO in a dose- and time-dependent manner by inducing apoptosis and necrosis of the tumor cells. The concurringly decreased glutathione peroxidase activity and the reduced mitochondrial membrane potentials may be possible underlying mechanisms for the apoptosis and necrosis of the tumor cells.
Wang
Chunfang
C
Beijing Institute of Ophthalmology, Beijing TongRen Eye Center, Beijing TongRen Hospital, Beijing, China.
Li
Bin
B
Zhang
Haiyan
H
Shi
Guangpu
G
Li
Weihong
W
Jonas
Jost B
JB
D017209:Apoptosis / Q000187:drug effects; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000008:administration & dosage / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D006131:Growth Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D008545:Melanoma / Q000201:enzymology / Q000473:pathology* / Q000503:physiopathology; D008564:Membrane Potentials / Q000187:drug effects; D008854:Microscopy, Electron; D008928:Mitochondria / Q000187:drug effects; D009336:Necrosis; D009994:Osmolar Concentration; D010087:Oxides / Q000008:administration & dosage / Q000494:pharmacology*
D001152:Arsenicals; D006131:Growth Inhibitors; D010087:Oxides; D005979:Glutathione Peroxidase; D000077237:Arsenic Trioxide