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目的 研究葡萄膜黑色素瘤细胞系中凝血酶敏感蛋白-1(thrombospondin-1,TSP-1)表达水平与葡萄膜黑色素瘤患者临床病理特征与肿瘤血管生成机制之间的关系.设计实验研究.研究对象葡萄膜黑色素瘤组织标本,葡萄膜黑色素瘤细胞系MUM-2B.方法 2004年1月至2007年12月在北京同仁医院行眼球摘除手术并经病理证实为葡萄膜黑色素瘤组织的标本50例,应用免疫组织化学法检测石蜡标本中TSP-1的表达,分析其与性别、年龄、组织学类型、肿瘤大小、侵犯睫状体、侵犯巩膜导管相关性.应用免疫组织化学法检测石蜡标本中CD34的表达,计算微血管密度(microvessel density,MVD),检测TSP-1与其相关性.构建TSP-1过表达慢病毒载体转染UM细胞系MUM-2eB,运用Q-PCR以及Western蛋白印迹法检测转染前后细胞系VEGF的表达量变化.主要指标葡萄膜黑色素瘤组织标本中TSP-1的表达与临床资料相关性,组织标本中MVD,细胞系TSP-1过表达前后VEGF表达量变化.结果 50例葡萄膜黑色素瘤组织中侵犯睫状体与未侵犯睫状体TSP-1表达阳性者分别为5/20和27/30;大肿瘤(最大基底直径＞16 mm或高度＞10 mm)和中等大小肿瘤(最大基底直径＜16 mm或高度在2.5～10 mm)表达阳性者分别为3/11和29/39(x2=-5.470、4.845,P＜0.05).不同性别、年龄、组织学类型以及是否侵犯巩膜导管间TSP-1表达阳性例数的比较差异均无统计学意义(P=0.664、0.606、0.856、0.723).葡萄膜黑色素瘤细胞系中TSP-1表达阳性组与阴性组的MVD差异具有统计学意义(Z=-4.616,P＜0.05).构建TSP-1过表达慢病毒载体转染UM细胞系MUM-2B,转染前后VEGF含量较前下降(P＜0.05).结论 TSP-1与葡萄膜黑色素瘤血管生成相关,葡萄膜黑色素瘤组织中TSP-1表达升高可抑制肿瘤血管生成,提示TSP-I可能作为预测葡萄膜黑色素瘤发生发展以及分子治疗靶点的指标.“,”Objective To investigate the relationship between TSP-1 expression and clinical pathological features of uveal melanoma patients and angiogenesis mechanism of uveal melanoma.Design Experimental study.Participants cell line MUM-2B and tissue specimen of uveal melanoma.Method 50 samples of uveal melanoma were collected in Beijing Tongren Hospital.The expression of TSP-1 and CD34 (microvessel density) in the samples was detected using immunohistochemistry.Building the overexpression of TSP-1 lentiviral vector and transfecting the uveal melanoma cell line MUM-2B,the variety expression of VEGF mRNA and protein was detected using Q-PCR and Western Blot.Main Outcome Measures The expression of TSP-1 and microvessel density in the uveal melanoma tissues,the clinical data,and the change of VEGF expression in the cell line before and after the TSP-1 overexpression.Results TSP-1 was positively expressed in 5 samples of 20 patients with ciliary body invasion and 27 samples of 30 patients without ciliary body invasion.There were 3 samples and 29 samples exhibiting the positive expression of TSP-1 in 11 large tumors and 39 medium tumors of uveal melanoma,which were respectively with the significant difference (x2-=5.470,4.845;P＜0.05).No significant differences in the expression of TSP-1 were found between different age,gender,cell type and sclera catheter (P=0.664,0.606,0.856,0.723).The significant differences the microvessel density were found between the positive and negative expression of TSP-1 (Z=-4.616,P＜0.05).Building the overexpression of TSP-1 lentiviral vector and transfecting the uveal melanoma cell line MUM-2B,the significant increasing were found in VEGF expression after transfection compared with before the transfecting.Conclusions TSP-1 expression is associated with the uveal melanoma angiogenesis mechanism.The increasing expression of TSP-1 can inhibit the angiogenesis of the tumor.These results imply that TSP-1 expression can be a molecular therapeutic target of uveal melanoma.