Objective To observe the macular morphological changes of choroidal melanoma with coherence tomography (OCT) after plaque radiotherapy (PRT). Methods A total of 48 patients (48 eyes) with choroidal melanoma who underwent125I PRT were enrolled in this study. All the patients were examined documenting OCT to get the image of macula. The macula of all the patients was not involved. The median visual acuity was 0.4plusmn;0.2, which ranged from 0.02 to 1.0. There were 18 eyes (37.5%) with retinal detachment, 12 eyes (25.0%) with retinal pigment epithelium (RPE) changes, seven eyes (14.6%) with macular edema, epimacular membrane, detachment combined with edema, exudation and RPE changes, 11 eyes (22.9%) with normal macular structure. The median follow-up time was (10.4plusmn;5.9) months, which ranged from one to 24 months. The tumor control situation and visual acuity were observed in follow-up period. The same equipment and methods of OCT were used to return visit in follow-up period. The macular morphological changes at the final visit and its relationship with PRT and visual acuity were contrastively analyzed. Results All the patients had good control of tumor. The vision acuity improved in two eyes (4.2%), unchanged in 10 eyes (20.8%), and decreased in 36 eyes (75.0%). The differences of the visual acuity was statistically significant between before and after treatment (Z=-3.778,P＜0.05). There were 13 eyes (27.1%) with retinal detachment; nine eyes (18.8%) with RPE changes; 17 eyes (35.4%) with macular edema, detachment combined with edema, exudation and RPE changes; six eyes (12.5%) with proliferation, atrophy, detachment combined with edema, exudation and epimacular membrane;three eyes (6.3%) with normal macular structure. There were 15 patients (31.3%) with two or more abnormal macular morphology after PRT. Conclusions Retinal detachment, RPE changes, macular edema and exudation are common abnormal macular morphology after PRT. The incidence rate of abnormal macular morphology is increased. There are 31.3% patients with two or more abnormal macular morphology.
Objective To observe the effects of immunologic cytokines or anti-angiogenesis gene transfer mediated by electroporation for choroidal melanoma cells.Methods The human embryo kidney cells and malignant choroidal melanoma cells were transfected with plasmids pNGVL-mIL2, pNGVL-mIL12, pCI-sFLK-1, pCR3.1-antiVEGF121,pCI-ExTek. Then the expression of mIL2, mIL12, sFLK-1, VEGF and ExTek were detected by enzymelinked immunosorbentassay (ELISA) and Western blot. Nude mice models of malignant choroidal melanoma were established and they were divided into four groups randomly. Each group was treated with 30 mu;l of 0.9% NaCl, 30 mu;g pNGVL, 30 mu;g antiVEGF121+sFLK-1+ExTek and 30 mu;g mIL2+mIL12 respectively by electroporation. Seven，14, 21, 28, 35 and 42 days after treatment, the tumor volumes were measured to calculate the tumor inhibition rate. Results ELISA and Western blot showed that mIL2，mIL12，sFLK-1 and ExTek were expressed after electroporation，VEGF expression was decreased remarkably. After treatment，the tumors of mIL2+mIL12 group were greatly inhibited with a tumor inhibition rate of 97.33%，while the tumors of antiVEGF121+sFLK-1+ExTek and pNGVL group were partially inhibited with tumor inhibition rates of 53.33% and 36.33% respectively.Conclusions Immunologic cytokines transfer mediated by electroporation can inhibit the growth of melanoma，but anti-angiogenesis only have a mild effects.
Objective To observe the therapeutic efficacy and complications of plaque radiotherapy (PRT) combined with transpupillary thermotherapy (TTT) on choroidal melanoma (CM). Methods Thirty unilateral CM patients (30 eyes, including 15 males and 15 females) were treated by PRT and TTT. The visual acuity ranged from 0.1 to 0.8 with an average of 0.3plusmn;0.2. The largest base diameter of tumor ranged from 6.8 mm to 17.9 mm with an average of (11.3plusmn;2.8) mm；The tumor height ranged from 3.9 mm to 10.6 mm with an average of (7.2plusmn;2.4) mm. The criteria of controlled local tumor: based on B-scan ultrasound measurement, the tumor was considered as ldquo;growingrdquo; if tumor height increased 2 mm or tumor largest base diameter increased 250 mu;m, otherwise the tumor was considered ldquo;controlledrdquo;. The followup ranged from 15 to 57 months with an average (33.01plusmn;9.81) months. The local tumor control rate, enucleation rate and visual acuity, complications after treatment were observed.Results The tumor largest base diameter after treatment ranged from 4.6 mm to 17.0 mm with an average (9.79plusmn;3.35) mm, which had statistically significant difference（t=2.195，F=0.49；P=0.032) with that before treatment. The tumor height after treatment ranged from 2.7 mm to 11.9 mm with an average (5.19plusmn;2.57) mm, which had statistically significant difference（t=2.069，F=0.018；P=0.0435) with that before treatment. At the end of follow up, the tumor largest diameter and height increased in two eyes respectively compared with those before treatment. Local tumor control rate was 86.7%. Three eyeballs were enucleated after treatment，the enucleation rate was 10.0%. The visual acuity remained unchanged in 12 eyes，improved in one eye and decreased in 17 eyes. Treatment complications included radiation retinopathy in 12 eyes (40.0%), secondary retinal detachment in three eyes (10.0%), secondary glaucoma in one eye (3.3%), cataract in four eyes (13.3%) and dry eye syndrome in five eyes (16.7%). Conclusion PRT combined with TTT is an effective therapy for choroidal melanoma with less complications.
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 influence of vascular endothelial growth factor (VEGF) antagonist bevacizumab on the growth of human choroidal melanoma (CM) OCM-1 cell xenografts in nude mice, and to explore the probable mechanism.Methods OCM-1 cells were subcutaneously implanted on 18 nude mice to establish ectopic model of human CM. The nude mice with the tumor of 5 mm in diameter were randomly divided into three groups: untreated group (group A), normal saline （NS） group (group B), drug treated group (group C). Bevacizumab was intraperitoneally injected for 14 consecutive days in group C, and the same volume of NS was used at a same way in group B. The volume and weight of implanted tumor as well as inhibitory rates of drug on tumor were calculated, ki67 and survivin proteins were measured with immunohistochemistry, and the mRNA expression of VEGF and survivin were assessed by RT-PCR.Results The volume and weight of tumor was (598.86plusmn;321.81) mm3, (0.66plusmn;0.15) g; (1 715.15plusmn;278.16) mm3, (1.54plusmn;0.39) g and (1 750.23plusmn;206.36) mm3, (1.54plusmn;0.31) g in groups C, A and B, respectively. There were significant differences between group C and A (F=34.53, P=0.00) and group C and group B (F=8.69, P=0.01). The inhibitory rate of these three groups were 57.14%, 5.31%, 6.25%, respectively, and the proliferation index (PI) of ki67 in these three groups were (51.85plusmn;1.32)%, (46.30plusmn;1.39)%, (27.90plusmn;0.90)%, respectively, there were significant differences in ki67 PI between C group and A or B group (H=15.17, P=0.00). The expression of survivin mRNA was (0.49plusmn;0.02), (0.82plusmn;0.05) and (0.61plusmn;0.05) in groupss C, A and B, respectively, there were significant differences between C group and A or B group (F=15.17, P＜0.05) . The expression of VEGF mRNA was (0.32plusmn;0.08), (0.73plusmn;0.07), (0.80plusmn;0.04) in groups C, A and B, significant difference was found between group C and A or B group (F=12.05，P＜0.05). Conclusion Bevacizumab can inhibit the growth of human CM in nude mice probably by inhibiting the activity of VEGF and downregulating survivin expression of the tumor as well as inhibiting the growth of the tumor.
Objective To investigate the expression of pigment epitheliumderived factor (PEDF) and vascular endothelial growth factor (VEGF) in choroidal melanoma. Methods The expression of VEGF and PEDF protein in fifty-eight cases of paraffinembeded choroidal melanoma samples was measured by immunohistochemistry, the expression of PEDF mRNA in thirtynine choroidal melanoma samples was assayed by in situ hybridization. Results PEDF protein was detected in 13/58 cases (22.4%) of choroidal melanoma, the positive rate in nonsclerainvasion group (12/38, 31.6%) was higher (Plt;0.05) than that in sclerainvasion group (1/20, 5%). VEGF protein was detected in 43/58 cases (64%) of choroidal melanoma, the positive rate in nonsclerainvasion group (25/38, 65.8%) was lower (Plt;0.05) than that in sclerainvasion group (18/20, 90%). The expression of PEDF mRNA was detected in 18/39(46.2) cases, the positive rate in nonsclerainvasion group was higher (Plt;0.05) than that in sclerainvasion group. Conclusions Imbalanced expression of VEGF and PEDF in choroidal melanoma may play a key role in the angiogenesis, tumor progression and metastasis.