Purpose: To characterize the safety profile of triamcinolone acetonide (TA) for intraocular application.Methods: In vitro cell viability assay was performed on 2 types of human ocular cells to evaluate the cytotoxicity of the simulated different vitreal concentrations (from a 1: 15 dilution as if injected into 1.5 mL of rabbit vitreous to 1: 50 dilution as if injected into 5 mL of human vitreous) of preservative and excipient (supernatant) from Kenalog-40. In vivo 35 guinea pigs were used for evaluating either a dose of intravitreal triamcinolone acetonide (Kenalog-40 and Triesence) or the supernatant of Kenalog-40. The animal eyes were monitored by biomicroscopy, ophthalmoscopy, tonometry, electroretinography, and histology.Results: A >= 1:15 dilution of triamcinolone acetonide supernatant from Kenalog-40 did not show cytotoxicity on cultured human pigment epithelial (retinal pigment epithelium) cells or Muller cells. In vivo, neither intravitreal 6 mu L (0.248 mg, equivalent to 4 mg in 0.1 mL for human eyes) nor 18 mu L (0.744 mg, equivalent to 4 mg in 0.1 mL for rabbit eyes and equivalent to 12 mg in 0.1 mL for human eyes) of triamcinolone acetonide suspension showed ocular toxicity. No significant difference was noted between Kenalog-40 and Triesence clinically and histopathologically.Conclusion: The equivalent triamcinolone acetonide doses to 0.1 mL (4 or 12 mg) intravitreal injection for human eye were found safe in guinea pig eyes. No significant difference was noted for 0.1 mL intravitreal injection between Kenalog-40 and Triesence. RETINA 32:364-374, 2012

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 μmol/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 μmol/L indomethacin (F= 16. 679, P＜0.01). The concentrations of survivin were (787.3±47.37), (257.0± 26.21), (123. 3± 8.02) pg/ml in control group and 100, 400 μmol/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.

Background Researches demonstrated that epidermal growth factor (EGF) can pmmote the proliferation and migration of Mailer cells of rat.However,whether EGF plays the role on human Müller cells is unknown.Objective Present study was to address if Müller ells express and secret EGF and explore the effects of EGF on the proliferation and migration of Müller cells under the normal and low oxygen conditions.Methods Human Müller cell line MIO-M1 cells were cultured and incubated in DMEM with high glucose and different concentrations of EGF in the presence or absence of varied amounts of GoCl2 for indicated time points.The preliferation and migration of Müller cells were analyzed with MTT and Transwell assay respectively.The expression and secretion of EGF in Müller cells were measured by using ELISA.Western blot was performed to detect and assess the protective role of ERKI／2 and Akt signal pathway under the presence of EGF.Results EGF stimulated the proliferation and migration of Müller cells in a concentration-dependent manner with a significantly different A570 values in the Müller cells among the various groups (F=123. 765, P = 0. 000). The maximum proliferation rate of Müller cells was found in 100 mg/L EGF group with the 1.6 times of elevation more than the free-EGF cells. Also,the 4. 5 times of increase was reached in migration rate in 10 mg/L EGF group more than free-EGF cells. In hypoxia condition,the median lethal concentration of CoCl2 was 600 nmoL/L. After pretreated with 10-100 mg/L EGF,the A570 values of Müller cells were obviously elevated with the statistical difference among various concentrations of EGF groups (F=22. 900 ,P=0. 000). The ELISA results showed that Müller cells secreted 7.8 μg/L EGF in the presence condition of 700 nmoL/L CoCl2. Western blot revealed that the presence of CoCl2 reduced the expression of ERK1/2 and Akt in Müller ells,however,pretreatment with EGF antagonized the harmful effect of CoCl2 on Müller cells.Conclusion EGF can induce the proliferation and migration of Müller cells at the dose-dependent pattern. Müller cells do not express and secrete EGF by themselves in normal condition, but hypoxia induce the expression and secretion of EGF. It is indicated that EGF promote the proliferation and migration of Müller cells through activating ERK1/2 and Akt signal pathways. External EGF can protect Müller cells against CoCl2 -induced cell damage.

PURPOSE:MicroRNAs (miRNAs) can contribute to tumorigenesis by acting as either oncogenes or tumor suppressor genes. The authors' previous studies on miR-34a showed that miRNA can influence the growth of uveal melanoma cells. In this study, they investigated the role of miR-137 in the pathogenesis of uveal melanoma.METHODS:Real-time RT-PCR was used to screen the expression levels of miR-137 in uveal melanocytes and uveal melanoma cell lines. Cell proliferation was examined by MTS assay and cell cycle was analyzed by flow cytometry. The target genes of miR-137 were predicted by bioinformatics and confirmed using a luciferase reporter assay. The expression of MITF, CDK6, and cell cycle regulatory proteins was determined by Western blot analysis. The ability to increase miR-137 expression by epigenetic drugs was tested using real-time RT-PCR.RESULTS:miR-137 expression was lower in uveal melanoma cell lines than in uveal melanocytes. Ectopic transfection of miR-137 into uveal melanoma cells induced G1 cell cycle arrest, leading to a significant decrease in cell growth. Overexpression of miR-137 downregulated MITF, a transcription factor with oncogenic activity. Moreover, the introduction of miR-137 downregulated the oncogenic tyrosine kinase protein receptor c-Met and cell cycle-related proteins, including CDK6. One avenue to increase the expression levels of miR-137 was through treatment with a DNA hypomethylating agent, 5-aza-2'-deoxycytidine, and a histone deacetylase inhibitor, trichostatin A.CONCLUSIONS:The results showed that miR-137 can act as a tumor suppressor in uveal melanoma cell proliferation through downregulation of the targets MITF and CDK6. miR-137 may be epigenetically silenced during uveal melanoma tumorigenesis.