ObjectiveTo observe the effectiveness of radiotherapy for refractory choroidal hemangioma. MethodsEight patients (8 eyes) with choroidal hemangioma were enrolled in this retrospective study. All the patients had received laser or photodynamic therapy before without effectiveness. The patients included 7 males and 1 females. The age was ranged from 11 to 54 years old, with an average of (27.50±15.18) years. All the patients were affected unilaterally, including 3 right eyes and 5 left eyes. There were 5 eyes with circumscribed choroidal hemangioma, 3 eyes with diffused choroidal hemangioma. All eyes had extensively exudative retinal detachment. The vision was from light sensation to 0.01. The volume of the tumors was ranged from 1.96 to 5.35 cm3, with a mean of (3.37±1.06) cm3. All the patients were treated with X rays by conventional fractional radiotherapy. Four of 8 patients were applied 24Gy totally in 8 fractions, while the other 4 patients were applied 46Gy in 23 fractions. Follow-up period ranged from 7 to 95 months, with medium of 42 months. ResultsRetinas reattached in all the eyes while exudation being absorbed. No retinal detachment happened again. To the last follow-up, the vision was from light sensation to 0.6. Visual activity improved in 6 eyes while 2 eyes improved obviously. Visual acuity was stable in remaining 2 eyes. The volume of the tumors decreased to 1.24-2.16 cm3, with a mean of (1.68±0.30) cm3. The percentage of the tumor decreased by 14.6-72.7, with an average of (44.89±21.30)%. No radiotherapy-associated complication occurred. ConclusionRadiotherapy is an efficient and safe treatment for refractory choroidal hemangioma.
Objective To isolate and purify the melanoma stem cells (MSC) in choroidal melanoma OCM-1 cells. Methods OCM-1 cells were resuscitated, and after cultured in standard Dubecco's modifided Eagle's medium (DMEM)/F12, they were cultured in serum-free medium (SFM). The cultured MSC were isolated and purified, and the positive rate of CD133, the specific markers of neurostem cells, was observed by flow cytometry (FCM). The 6th generation of the cells were stained by musashi-1 immunocytochemistry, and the rate of the positive cells was observed under the microscope. Results After the Adherent OCM-1 cells cultured in SFM, the number of the adherent number decreased obviously. The cells at the 6th generation grew as the suspended gobbets, which represented the typical grow manner of the stem cells. Positive CD133 could be found in the cells of different generations, which was 2.5％, 21.7％, and 57.8％ in the non-isolated OCM-1 cells, the 1st generation of isolated cells, and the 2nd generation cells, respectively. The positive rate of CD133 in the cells at the sixth generation was 79.8% with b positive expression of musashi-1. Conclusion MSC is in the human choroidal melanoma OCM-1 cells. The suspended stem cells may be purified by limited differentiation and serial passage in SFM. (Chin J Ocul Fundus Dis, 2007, 23: 87-90)
ObjectiveTo observe the expression and transcription of MART-1 in human uveal melanoma cell lines 92-1, 92-2, Ocm3, Me1285, as well as the possible effect of methylation on its expression.MethodsThe cell lines 92-1, 92-2, Ocm3 and Mel285 were cultured routinely and tested for MART-1 expression at protein and mRNA level by FACS analysis, Western blot and RT-PCR respectively. Methylation status of the MART-1 promoter region in all the cell lines were checked by Southern blots of DNA digested with methylation sensitive restriction enzymes.ResultsAs observed in FACS analysis and Western blot, 92-1, 92-2 and Ocm3 were MART-1 positive cell lines while Me1285 was negative cell line. Consistent with protein analysis, 92-1 and Ocm3 cell lines showed MART-1 specific PCR products and there was no product in Me1285 cell line in RT-PCR. The MART-1 positive cell lines, 92-1, 92-2, and Ocm3 show methylation at the MspI/HpaⅡ site, and the NruⅠ sites of all positive cell lines are not methylated. The MART-1 negative cell line Mel285 shows hypermethylation at the NruⅠsite and the MspⅠ/HpaⅡ site is not methylated.ConclusionsMART-1 could be expressed in human uveal melanoma cell lines 92-1, 92-2 and Ocm3. The change of methylation status of MART-1 promoter may correlate with the transcription of MART-1.
ObjectiveTo observe the OCT angiography (OCTA) imaging features of isolated choroidal hemangioma (CCH).MethodsA retrospective case study. From January 2017 to February 2019, 18 CCH patients (18 eyes) diagnosed in the Affiliated Eye Hospital of Nanjing Medical University were included in the study. There were 13 males (13 eyes) and 5 females (5 eyes), with the mean age of 44.5 years. All the tumors were orange-red, with clear boundaries, located at the posterior pole or around the optic disc. OCTA was used to scan the 6 mm×6 mm of macular area or in the range of 6 mm×6 mm. After automatic image processing, the system provided the blood flow map of shallow capillary plexus, deep capillary plexus, outer retina and choroidal capillary plexus, as well as the corresponding structure en-face image and B-scan image.ResultsOCTA examination found that when the stratification line was adjusted to the periphery of the choroidal capillary layer, the blood flow map showed clear boundary of the tumor, and the blood vessels on the surface of the tumor presented a network crisscross with different thickness. B-scan image showed that the whole layer of retinal choroid at the tumor presented a dome-shaped uplift, and the neurocortical layer could be accompanied by thickening, subretinal effusion, exudation and splitting. En-face image showed that the boundary of the tumor was clear, the surrounding exudation was strong reflection in spots or patches, local pigmentation showed weak reflection, and the signal reflection was uneven.ConclusionOCTA can clearly show the vascular morphology on the surface of CCH.
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.