Objective To investigate the development and metastasis of malignant choroidal melanoma cell strain OCM-1-gfp modified with green fluorescent protein(GFP) and the factors which affected the tumor biological behaviors. Methods GFP was transfected into malignant melanoma cell strain OCM-1.Melanoma cells with high and stable expression of GFP were injected into subretinal space and the subcutaneous space of hind leg of Balb/c nude mouse respectively in order to establish orthotopic and heterotopic transplanted tumor models.The development and metastasis process of orthotopic tumor models was observed directly by fluorescence microscope，and the size of the hypodermal tumor was measured by vernier.The expressions of 13 genes in melanoma were detected by means of immunohistochemistry staining. Results Malignant choroidal melanoma cell strain OCM-1 stably expressed GFP and preserved the characteristics of parental generation，OCM-1-gfp may develop melanoma and continue to metastasize in nude mouse.Positive expression of most of the antibodies，including Rb，p53，p21，E2F，NFkappa;B，cyclin D1，proliferation cellular nuclear antigen（PCNA），bcl2、bclXL/S，bax，and epithelial growth factor（EGF）and its receptor（EGFR），was found.While the staining of inhibition gene p16 was negative. Conclusions GFP is the marker for observing the development and metastasis of malignant choroidal melanoma in vivo.The rate of tumor formation and development process in orthotopic models does not differs much from which in heterotopic models of malignant choroidal melanoma.The expressions of lots of genes in malignant choroidal melanoma developed from OCM-1-gfp including p16、p53、NFkappa;B，cyclin D，PCNA，EGF，and EGFR are abnormal. (Chin J Ocul Fundus Dis, 2006, 22: 170-173)
Objective To investigate the histopathological characteristics of choroidal melanoma. Methods The histopathological data from 64 patients with choroidal melanoma were analyzed retrospectively. The tumor size and the cytological types were observed and detected. The locations of the tumor were classified according to the involved part invaded by the anterior margine of the tumor, and the degrees of the development of the tumor were graded according to the extent of the outward infiltration of the tumor cells. Results In 64 patients with choroidal melanoma, There were large, medium, and small tumors in 25(39.1%), 31(48.4%), and 8(12.5%) respectively. The spindle cell type was found in 42 patients (65.6%) including spindle cell A and B type in 15(23.4%) and 27(42.3%) respectively; epithelioid and mixed cell type was found in 7(10.9%) and 10(15.6%) respectively; the other types were found in 5(7.8%). Twenty-five cases（39.1％）had no invasion with sclera, 22（34.4％）had but limited to sclera, 12（18.8％）penetrated through sclera and 5（7.8％）had intra-orbit infiltration. Conclusion The histopathological characteristics of choroidal melanoma are multiple, and spindle cell type is the most common one. The choroidal melanoma can easily invade the sclera. (Chin J Ocul Fundus Dis, 2006, 22: 161-165)
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)
Objective To evaluate the clinical and histopathological features of diffuse choroidal melanoma. Methods The clinical and histopathological data of 11 patients with diffuse choroidal melanoma were reviewed retrospectively. Those patients were referred to Tianjin Eye Hospital because of visual loss or ophthalmalgia (10 cases), or Coats disease with secondary glaucoma and atrophy bulbi (1 case). The clinical disgnosis included choroidal tumor or melanoma (8 cases), absolutestage glaucoma (2 cases) and atrop hy bulbi with Coats disease (1 case). Nine patients received enucleation, and 2 patients received enucleation combined with orbital exenteration. The cellular proliferation was assessed by Ki-67staining. Results All 11 tumors had grown flatly with a wide base ranged from 12 to 20 mm, and tumor thickness ranged from 2 to 4 mm. There were 9 cases of mixed cell type, 1 case of epithelioid cell type and 1 case of necrotic cell type. The tumors invaded into the sclera in 7 cases and orbital cavity in 3 cases. Secondary glaucoma was found in 7 cases. On average, 9% (7%13%) of tumor cells were Ki67 positive and most of them located at the tumor base. There were more Ki67 positive epithelioid tumor cells than Ki67 positive spindle-shaped cells. Conclusions Diffuse choroidal melanoma had a special growth pattern and is difficult to be recognized, sometimes could be misdiagnosed as glaucoma or other choroidal tumors. With its wide base, this tumor could easily invade the orbit and metastate, and its prognosis is very poor.