葡萄膜黑色素瘤染色体异常及其相关基因研究进展

葡萄膜黑色素瘤存在多种染色体异常及基因表达异常。3号染色体缺失和8号染色体获得与葡萄膜黑色素瘤的转移和预后密切相关，可作为其分型和预后评判的指标；6号染色体的异常与葡萄膜黑色素瘤不同表型的侵袭能力有关。Cyclin D1、MTS1、MDM2、Bcl-2等基因表达异常与葡萄膜黑色素瘤的发生有关，DDEF1、NBS1基因的表达与葡萄膜黑色素瘤的侵袭能力和转移密切相关。     

葡萄膜黑色素瘤转移相关基因的表达及意义

目的：探讨肿瘤转移相关基因在葡萄膜黑色瘤中的表达及与浸润能力和病理分型的关系。方法：采用免疫组化方法定理检测96例葡萄膜黑色素瘤体标本中EGFR和nm23蛋白的表达，结果：肿瘤的浸润能力与EGFR的表达呈正相关，与nm23的表达则呈负相关，类上皮细胞型，混合细胞型，梭形细胞型EGFR的表达率依次降低，nm23的表达率依次增高，结论：EGFR和nm23是评价葡萄膜黑色素瘤转移潜能的有效指标。     

E26转录因子-1在不同类型葡萄膜黑色素瘤中的表达及意义

目的　探讨E2 6转录因子 1(E2 6transformation specific 1,Ets 1)在不同类型的葡萄膜黑色素瘤中的表达及其与预后的相关关系。方法　以原位杂交和免疫组织化学法检测 78例葡萄膜黑色素瘤中Ets 1mRNA和蛋白的表达 ,并按WHO 1980年的标准进行分型 :梭型细胞型、类上皮细胞型和混合细胞型。结果　 78例中 ,梭型细胞型占 2 1例 ,类上皮细胞型占 34例 ,混合细胞型占 2 3例。Ets 1mRNA和蛋白在 3种类型的葡萄膜黑色素瘤中均有表达 ,但表达强度随梭型细胞型 ,混合细胞型和类上皮细胞型依次递增。回访37例患者 ,其中梭型细胞型 18例 ,平均生存时间为 (78.33± 2 4 .6 9)月 ;混合细胞型 10例 ,平均生存时间 (6 1.4 4± 2 0 .4 6 )月 ;类上皮细胞型 9例 ,平均生存时间 (36 .76± 12 .19)月。患者生存时间与Ets 1表达强度呈负相关。结论　Ets 1可能在葡萄膜黑色素瘤的转移、浸润中发挥重要作用 ,Ets 1的检测可作为葡萄膜黑色素瘤恶性程度的参考指标。     

转化生长因子-β1在葡萄膜黑色素瘤中的表达及意义

目的:探讨转化生长因子-β(1TGF-β1)在葡萄膜黑色素瘤中的表达及意义。方法:以原位杂交法检测78例葡萄膜黑色素瘤中TGF-β1mRNA的表达,并按WHO1980年的标准进行分型:梭型细胞型,类上皮细胞型和混合细胞型。进行临床随访。结果:78例中,梭型细胞型21例,类上皮细胞型34例,混合细胞型23例。TGF-β1mRNA在3种类型的葡萄膜黑色素瘤中均有不同程度的表达,表达强度随梭型细胞型,混合细胞型和类上皮细胞型依次递增。回访37例患者,其中梭型细胞型18例,平均生存时间为(78.3±14.2)mo,混合细胞型10例,平均生存时间(69.1±17.4)mo,类上皮细胞型9例,平均生存时间(36.8±12.2)mo。患者生存时间与TGF-β1表达强度呈负相关。结论:TGF-β1可能在葡萄膜黑色素瘤的转移,浸润中发挥重要作用。     

血清黑色素瘤活性抑制蛋白与葡萄膜黑色素瘤

目的检测葡萄膜黑色素瘤患者血清黑色素瘤活性抑制蛋白(MIA)的水平，并探讨其在葡萄膜黑色素瘤诊断和转移监测中的价值。方法应用酶联免疫吸附法（ELISA）分别检测不同组织病理分型的葡萄膜黑色素瘤（27例）、葡萄膜黑色素细胞瘤（6例）、其他眼部肿瘤患者（7例）以及正常成人（16人）外周血清中MIA的浓度。结果正常成人(16人)和睫状体无色素上皮腺瘤(4例)、视网膜母细胞瘤(2例)、视网膜血管瘤(1例)患者血清MIA浓度显著低于葡萄膜黑色素瘤患者；不伴巩膜浸润和远处转移的葡萄膜黑色素瘤患者血清MIA浓度明显低于伴有巩膜浸润和远处转移的患者，但与葡萄膜黑色素细胞瘤患者比较无明显差异；在不伴巩膜浸润和远处转移的葡萄膜黑色素瘤组，梭形细胞型患者血清MIA浓度与混合型和上皮细胞型患者相比无明显差异。结论血清MIA水平可能是临床诊断葡萄膜黑素瘤的一个较好指标，并且可用于肿瘤转移的监测。(中华眼底病杂志,2005,21:153-155)     

葡萄膜黑色素瘤患者血清血管内皮生长因子浓度及临床意义

目的测定葡萄膜黑色素瘤患者血清血管内皮生长因子(VEGF)的水平,并探讨其临床意义。方法应用酶联免疫吸附法(ELISA),分别检测不同组织病理分型的葡萄膜黑色素瘤(27例)、葡萄膜黑色素细胞瘤患者(6例)以及正常人(16例)外周血清中VEGF的质量浓度。结果正常成人和葡萄膜黑色素细胞瘤患者血清VEGF浓度显著低于葡萄膜黑色素瘤患者;有巩膜导水管浸润和远处转移的葡萄膜黑色素瘤患者血清VEGF质量浓度明显高于不伴巩膜导水管浸润和远处转移的患者;梭形细胞型患者血清VEGF浓度与混合型和上皮细胞型患者经统计分析无显著差异。结论血清VEGF水平在监测葡萄膜黑色素瘤的转移中有一定的意义。     

反义bcl-2寡核苷酸对体外培养的葡萄膜黑色素瘤细胞多药耐药性的影响

目的：探讨体外培养的葡萄膜黑色素瘤细胞对不同化疗药物的敏感性,并通过反义bcl-2寡核苷酸（bcl-2 antisense oligodeoxynucleotide,bcl-2 AS-ODN）特异阻断bcl-2基因的表达逆转肿瘤耐药性。方法：原代培养葡萄膜黑色素瘤细胞,采用噻唑蓝染色法测定肿瘤细胞对不同浓度的5-氟尿嘧啶、噻替哌、顺铂、阿霉素、长春新碱和氮烯咪胺体外敏感性;通过阳离子脂质体导入bcl-2 AS-ODN,阻断bcl-2基因的表达,利用免疫组化及Western blot法测定肿瘤细胞bcl-2的表达情况,并根据多药相互作用原理测定肿瘤细胞对化疗药物敏感性。结果：葡萄膜黑色素瘤对不同种类的化疗药物均有不同程度的抗药性。Bcl-2 AS-ODN可明显抑制bcl-2基因的表达,并随浓度升高抑制作用增强。Bcl-2AS-ODN与各种化疗药物呈协同作用,能够增加化疗药物对肿瘤细胞的杀伤作用。结论：所选6种化疗药物在临床常用剂量范围对葡萄膜黑色素瘤细胞毒性作用较小,葡萄膜黑色素瘤细胞的多药耐药性与bcl-2基因的高表达有关,bcl-2 AS-ODN能够部分逆转肿瘤细胞的多药耐药性。     

间α胰蛋白酶抑制物在脉络膜黑色素瘤中的表达及意义

目的 探讨间α胰蛋白酶抑制物(ITI)在脉络膜黑色素瘤中的表达及与病理学分型和恶性程度、浸润能力的关系。方法 选择石蜡包埋脉络膜黑色素瘤组织28例，通过脱色素免疫组织化学方法和计算机图像分析技术进行ITI表达的检测。结果 ITI在瘤细胞胞浆中被检测到。梭形细胞型、混合细胞型、上皮细胞型ITI的阳性表达率依次降低。脉络膜黑色素瘤的恶性程度、浸润能力与ITI的表达呈负相关。平均吸光度值在各型脉络膜黑色素瘤中无明显不同。结论 脉络膜黑色素瘤表达ITI。其存在抑制了脉络膜黑色素瘤的恶化浸润和转移，且一定程度上可以反映其恶性程度及预后。     

比较蛋白质组学技术筛选葡萄膜黑色素瘤相关蛋白

目的 应用比较蛋白质组学技术,分析鉴定葡萄膜黑色素瘤与正常人葡萄膜差异蛋白质,筛选出葡萄膜黑色素瘤相关蛋白.设计实验性研究.研究对象手术中切除的人葡萄膜黑色素瘤标本4例以及正常人捐献眼葡萄膜标本4例.方法 研磨组织提取蛋白质后,应用双向凝胶电泳进行总蛋白分离,考马斯亮兰染色,Image Master 5.0软件分析蛋白胶图,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定部分差异蛋白质点,通过MASCOT软件在NCBI数据库中检索蛋白质信息.主要指标差异蛋白质.结果 建立了葡萄膜黑色素瘤和正常人葡萄膜组织的双向凝胶电泳图谱,鉴定了葡萄膜黑色素瘤与正常人葡萄膜之间差异表达的蛋白质30种,其中24种蛋白只在葡萄膜黑色素瘤中出现.葡萄膜黑色素瘤中表达上调的蛋白5种,表达下调的蛋白1种.这些差异蛋白可能执行新陈代谢酶、信号转导调节、细胞骨架、免疫调节等生物学功能.结论 通过比较蛋白质组学初步筛选出与葡萄膜黑色素瘤发生演进相关的蛋白质.需要进一步验证蛋白的定位及功能.(眼科,2008,17:230-234)     

E26转录因子-1、基质金属蛋白酶-1及其抑制剂在脉络膜黑色素瘤中的表达及意义

目的 探讨E26转录因子-1（E26ts-1）、基质金属蛋白酶-1（MMP-1）及其抑制剂（TIMP-1）在脉络膜黑色素瘤中的表达及其与肿瘤浸润及转移的关系。 方法 以免疫组织化学法检测78例葡萄膜黑色素瘤患者中的E26ts-1，MMP-1和TIMP-1的表达，并按肿瘤细胞的形态进行分型:梭型细胞型，类上皮细胞型和混合细胞型。进行临床随访，计算平均生存时间，以SPSS 10.0统计软件包进行统计学处理。 结果 78例脉络膜黑色素瘤患者中，梭型细胞型21例，类上皮细胞型34例，混合细胞占23例。TIMP-1呈低表达，E26Ts-1和MMP-1在三种类型的脉络膜黑色素瘤中均有表达;表达强度为梭型细胞型、混合细胞型和类上皮细胞型依次递增。共回访37例患者，其中梭型细胞型18例，平均生存时间为（78.33±24.69）个月;混合细胞型10例，平均生存时间（61.44±20.46）个月;类上皮细胞型9例，平均生存时间（36.76±12.19）个月。患者生存时间与E26ts-1及MMP-1的表达强度呈负相关（P＜0.01）。 结论 E26Ts-1和MMP-1的高表达及TIMP-1的低表达可能与脉络膜黑色素瘤的转移浸润有关。 (中华眼底病杂志, 20063, 22:174-176)     

Transporter associated protein expression in uveal melanoma

BACKGROUND/AIM: Transporter associated protein (TAP) is important for presenting peptides to major histocompatibility complex (MHC) class I molecules. Defects in TAP lead to decreased MHC class I expression. The immunoexpression of human leucocyte antigen (HLA) class I molecules and the TAP1 subunit were studied in primary uveal melanomas and correlated with the cell types and extrascleral extension. METHODS: The HLA class I antigen and TAP1 subunit were analysed by immunoperoxidase staining with monoclonal antibodies on 45 primary uveal melanoma archival specimens. The tumours were divided into two groups-group A: tumours with no extrascleral extension; group B tumours with extrascleral extension/liver metastasis. Immunoanalysis was done by a semiquantitative method. RESULTS: HLA class I antigen and TAP1 were decreased in 35 of 35 tumours with no extrascleral extension and positive in six of 10 tumours with liver metastasis. Decreased immunoexpression of HLA class I antigen and TAP1 in uveal melanomas with no extrascleral extension was significant (p<0.001). HLA class I antigen and TAP1 were negative in spindle cell melanomas (p<0.001). CONCLUSIONS: HLA class I antigen and TAP1 expression were decreased in uveal melanomas with no extrascleral extension and in spindle cell melanomas. Decreased expression of TAP1 may lead to decreased expression of HLA class I antigen in uveal melanomas. This preliminary observation deserves further investigation, which may shed more light on the immune escape mechanisms of this tumour and thus enable novel therapeutic strategies.     

Cripto-1 expression in uveal melanoma: an immunohistochemical study

Human Cripto, the founder member of the epidermal growth factor-Cripto-FRL1-Cryptic (EGF-CFC) family, plays an important role during early embryonic development and in particular in carcinogenesis and the development of cancer metastases. Cripto-1 is over-expressed in most cancers, but is absent or only weakly expressed in normal cells. For this reason, Cripto-1 could be of potential value in the targeted treatment. There is no information on the expression of Cripto-1 in human uveal melanoma. Cripto-1 reactivity was evaluated by immunohistochemistry on 36 archival uveal melanomas using the polyclonal antibody to Cripto-1. The tumors were divided in to 2 groups. There were 18 uveal melanomas with no intrascleral or extrascleral extension and 18 uveal melanomas with intrascleral/extrascleral extension/liver metastasis. Cripto-1 reactivity was correlated with tumor aggressiveness and cell type. Furthermore, we studied the immunolocalization of Cripto-1 in 4 uveal melanoma cell lines OCM-1, OCM-8, and 92-1, and OMM-1 and in 2 primary uveal melanocyte cultures. Cripto-1 was expressed in both the non-invasive and aggressive uveal melanomas. Cripto-1 was positive in the 4 uveal melanoma cell lines and absent in the primary uveal melanocyte cultures. Retinal tissue did not express Cripto-1. The results suggest that Cripto-1 is expressed in uveal melanoma, negative in the non-neoplastic ocular tissue and point to its use as a target for therapy.     

HLA expression in choroidal melanomas: correlation with clinicopathological features

PURPOSE: In this retrospective study, we analyzed the relevance of human leukocyte antigen (HLA) expression to clinical behavior of uveal melanoma and correlated it with conventional light microscopic parameters. METHODS: HLA class I antigen and Beta 2 microglobulin expression were analyzed in 45 primary choroidal melanoma lesions by immunoperoxidase staining with monoclonal antibodies to HLA class I antigen and beta 2 microglobulin and correlated with the known clinicopathological parameters. Immunoanalysis was done by a semi quantitative method. The tumors were divided into 3 groups. Group A: Tumors with no extrascleral extension/no liver metastases, group B: tumors with only extrascleral extension and group C: tumors with liver metastases. For statistical analysis we analyzed the negative, weak (heterogeneous) and the positive expression of HLA and beta 2 microglobulin with known clinicopathological parameters. RESULTS: In-group A (n = 35) tumors with no extrascleral extension and no liver metastases HLA class I antigen and beta 2 microglobulin are negative (absent staining) in 30 choroidal melanomas. In-group B (n = 4) they are weak (heterogeneous) in 3 tumors. In-group C (n = 6) all the 6 tumors have a positive (bright staining) immunoexpression. No statistical significance was obtained when HLA and beta 2 microglobulin immunoreactivity was compared against largest tumor diameter (LTD), tumor infiltrating lymphocytes (TIL), mitosis and nuclear grade. The difference of HLA class I and beta 2-microglobulin imunoreactivity among the various cell types spindle, mixed and epithelioid was statistically significant (p = 0.003), (p = 0.004). The difference in immunoreactivity between tumors with no liver metastases and tumors with liver metastases was statistically significant (p < 0.001). CONCLUSIONS: HLA class I antigen and beta 2 microglobulin are negative in melanomas with no extrascleral extension and liver metastases and weak in melanomas with extrascleral extension and are positive in melanomas with liver metastases. HLA expression is independent of the conventional markers in uveal melanoma.     

Orbital recurrence of choroidal melanoma 20 years after enucleation

A 54-year-old woman developed an orbital mass 20 years after enucleation of the left eye for a choroidal melanoma, which proved to be of spindle-A type. Computed tomography of the orbit suggested the diagnosis of a cavernous hemangioma, but orbital biopsy confirmed the diagnosis of recurrent malignant melanoma and the patient was managed by orbital exenteration followed by radiotherapy. The orbital recurrence was of the mixed cell type, suggesting a gradual 20-year transition of the spindle-A cells to spindle-B and epithelioid cells. We recommend that the selected surgical management of extrascleral extension of uveal melanomas be determined by the degree of extrascleral involvement and by whether the extrascleral extension is discovered clinically, at surgery, or in the pathology laboratory. We recommend that orbital computed tomography be performed periodically after enucleation in all patients who have orbital extension of uveal melanomas.     

Expression of epidermal growth factor receptor, ezrin, hepatocyte growth factor, and c-Met in uveal melanoma: an immunohistochemical study

The immunoreactivity of epidermal growth factor receptor (EGFR) ezrin, hepatocyte growth factor receptor (HGF), and c-Met was studied in 60 uveal melanomas and was correlated with clinicopathologic parameters. Metastases were diagnosed in the patients with uveal melanoma between 5 years and 8 years (median, 6.5 years) after enucleation. Using Kaplan-Meier statistical analysis, we found a significant association between high c-Met expression and death due to uveal melanoma (p < 0.03). EGFR was expressed in 18 of 60 (30%) tumors; ezrin was expressed in 30 of 60 (50%) tumors. Tumors with liver metastasis (n = 6) showed higher expression of c-Met (p = 0.0009) compared with the tumors with no extension/extrascleral extension without liver metastasis (groups A-45 and B-9). HGF was negative in all the six tumors that had liver metastasis. Further studies are required to understand the possible mechanism of ligand-independent c-Met activation in patients with uveal melanoma.     

Two discrete uveal melanomas in a child with ocular melanocytosis

OBJECTIVE: To describe a case of two uveal melanomas in a child with mild ocular melanocytosis. METHODS: A 6-year-old girl was followed for 5 years with an ill-defined, slowly enlarging presumed choroidal nevus in the postequatorial fundus. Ocular oncology evaluation revealed mild sectorial scleral and uveal melanocytosis and an episcleral sentinel vessel superotemporally. Two discrete uveal melanomas were present. In the circumpapillary and macular region, tumor 1 was diffuse at 9.0 mm in base and 4.1 mm in thickness and with overlying subretinal fluid. In the ciliary body, tumor 2 was discovered by transillumination and was 6.0 mm in base and 2.2 mm in thickness. Enucleation was performed. RESULTS: Histopathologic analysis disclosed two discrete uveal melanomas in a bed of diffuse mild uveal melanocytosis. Tumor 1 was a mixed, predominantly epithelioid cell melanoma with active mitotic figures, and tumor 2 was a mixed, predominantly spindle cell melanoma. The choroid between the melanomas showed only benign, dendritic melanocytes consistent with melanocytosis. There was no extrascleral extension. CONCLUSIONS: Ocular melanocytosis can predispose to one or multiple uveal melanomas. Lifetime ophthalmic monitoring of affected patients is warranted.     

Case report: ciliary body melanoma with extrascleral extension.

Uveal melanoma is an uncommon cancer that is most often diagnosed in the sixth decade and is somewhat more common in males. Early detection and treatment of melanoma is important as metastasis can be fatal. A case of ciliary body melanoma that was detected in an asymptomatic patient because of extrascleral extension is presented. Signs and symptoms that may lead to the diagnosis of uveal melanoma are presented, as well as lesions that may simulate extrascleral extension of melanoma.