Objective To observe the influence of the indomethacin on the proliferative and invasive activity of OCM-1 human choroidal melanoma cells. Methods OCM-1 cells were cultured with different concentrations of indomethacin (25, 50, 100, 200, 400 mu;mol/L ), and their proliferation were assessed by methyl thiazolyl tetrazolium（MTT）, invasive behaviors were examined by cell invasion assays, expression of survivin and VEGF were evaluated by reverse transcriptase polymerase chain reaction（RT-PCR）, immunofluorescence staining, ELISA and western blot analysis. Result All concentrations of indomethacin in this study can inhibit the proliferation and invasion of OCM-1 cells in a time and dosage-dependant manner（MTT/24 h：F=19.642，P＜0.01；MTT/48 h：F=136.597，P＜0.01；MTT/72 h：F=582.543，P＜0.01；invasion assays：F=54.225，P＜0.01）. Immunofluorescence staining indicated that survivin and VEGF mainly expressed in the cytoplasm of OCM-1 cells. Survivin mRNA in OCM-1 cells was inhibited by 100, 200, 400 mu;mol/L indomethacin（F=16.679，P＜0.01）. The concentrations of survivin were (787.3plusmn;47.37), (257.0plusmn;26.21), (123.3plusmn;8.02) pg/ml in control group and 100, 400 mu;mol/L indomethacin groups, respectively. Survivin expression was also significantly down-regulated in indomethacin-treated cells by Western blot analysis.Indomethacin had no effects on VEGF expression in OCM-1 cells.Conclusions Indomethacin can inhibit proliferation and invasion of OCM-1 cells in vitro,down-regulated expression of survivin may be the mechanism.
Objective To investigate the protective effect of nerve growth factor (NGF) on apoptosis of cultured human fetal retinal pigment epithelium (HFRPE) cells induced by indomethacin (IN) in vitro.Methods Subcultured HFRPE cells were treated with different concentrations of IN to establish apoptotic model. The protective effect of NGF on apoptosis of cultured HFRPE cells were assessed using an acridine orange (AO) staining method and transmission electron microscopy (TEM).Results HFRPE cells exposed by 200-600 μmol/L IN for 24 hours elicited typical apoptosis morphological changes, including condensed chromation, nuclear fragmentation and reduction of nuclear size and cell volume. There was a statistically difference in HFRPE cells with apoptosis between 200 μmol/L IN+500 μg/L NGF and 200 μmol/LIN groups ( q=3.9204，P=0.0320); there was a significant difference in HFRPE cells with apoptosis in 400 μmol/L IN+500 μg/L NGF and 400 μmol/ L IN as well (q=9.7915，P=0.0001). Conclusion NGF has an protective effect on IN-induced HFRPE cells apoptosis. (Chin J Ocul Fundus Dis，2003，19：38-41)