血管生成相关因子在脉络膜黑色素瘤中的表达

摘要

背景 血管生成与肿瘤的发生之间有着密切的关系,目前对脉络膜黑色素瘤的发生发展过程与缺氧及血管生成相关因子相互关系的研究尚属少见.目的 研究血管生成相关因子缺氧诱导因子1α(HIF-1α)、诱导型一氧化氮合酶(iNOS)、环氧合酶2(COX-2)及血管内皮生长因子(VEGF)在脉络膜黑色素瘤中的表达及其相互之间的关系.方法 收集青岛大学医学院附属医院眼科病理室保存的脉络膜黑色素瘤标本44例和眼睑色素痣石蜡标本16例,应用兔抗人HIF-1α、iNOS、COX-2及VEGF多克隆抗体,采用免疫组织化学SP法测定HIF-1α、iNOS、COX-2及VEGF在脉络膜黑色素瘤组织及眼睑色素痣中的表达并进行比较,评价4种蛋白与脉络膜黑色素瘤临床病例特征的关系,并对4种蛋白在脉络膜黑色素瘤中表达的相互关系进行研究.结果 脉络膜黑色素瘤组织中VEGF、HIF-1α、iNOS、COX-2的阳性表达率均明显高于眼睑色素痣组织,差异均有统计学意义(x2 =6.580、4.105、8.350、13.240,P＜0.05);VEGF蛋白在较大体积肿瘤中的表达明显高于体积较小的肿瘤,差异有统计学意义(x2=9.389,P＜0.05),但VEGF蛋白在不同病理分型及不同浸润深度肿瘤中的表达差异均无统计学意义(x2=1.186、5.398,P＞0.05)；HIF-1α在脉络膜黑色素组织中的表达与肿瘤体积大小、病理分型有关,差异均有统计学意义(x=8.664、6.622,P＜0.05),但其与肿瘤的浸润深度无关,差异无统计学意义(x2=4.914,P＞0.05)；iNOS蛋白的表达与肿瘤体积大小有关(x2=8.609,P＜0.05),但与病理类型及浸润深度无关,差异均无统计学意义(x2=4.789、4.309,P＞0.05)；COX-2蛋白的表达在不同体积的肿瘤中明显不同,差异有统计学意义(x2=7.320,P＜0.05),而COX-2蛋白在不同病理分型及浸润深度的肿瘤组织中的表达量差异均无统计学意义(x2=2.772、2.103,P＞0.05).HIF-1α、iNOS、COX-2的表达量与VEGF表达均呈显著正相关(r=0.943、1.000、0.986,P＜0.05)；COX-2与iNOS表达量之间、HIF-1α与iNOS表达量间及HIF-1α与COX-2的表达量间均呈明显正相关(r=0.986、0.943、0.986,P＜0.05).结论 HIF-1α、iNOS、COX-2在脉络膜黑色素瘤中表达升高,其表达水平与VEGF相关,提示3种基因对脉络膜黑色素瘤的血管生成均有一定作用,可通过该作用促进肿瘤组织的生长.%Background Angiogenesis is an essential event for the growth and mobility of malignant tumors,but there is little literature about the effects of anoxia and angiogenesis-related factors on angiogenesis in choroidal melanoma.Objective The aim of this study was to investigate the expression of angiogeneic factors such as hypoxia inducible factor-1α(HIF-1α),inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),and vascular endothelial growth factor(VEGF)in choroidal melanoma and their clinical significance.Methods The specimens from 44 cases of choroidal melanoma and 16 cases of eyelid nevi identified by pathology were collected.Immunohistochemistry SP was used to examine the expression of HIF-1α,iNOS,COX-2 and VEGF in the samples.The expression of HIF-1 α,iNOS,COX-2 and VEGF in tumors with different sizes,different pathological types and different growth patterns was evaluated.The correlations among these four proteins in tumor growth and development were assessed.Results The expression of HIF-1α,iNOS,COX-2 and VEGF in choroidal melanoma were significantly higher than those in the eyelid nevi group(x2 =6.58,4.105,8.350,13.240,P＜0.05).The expression of VEGF was associated with tumor size(x2 =9.389,P＜0.05),but not associated with pathological types(x2 =1.186,P＞0.05)and infiltration depth(x2 =5.398,P＞ 0.05).The expression of HIF-1α was associated with tumor size(x2=8.664,P＜0.05)and pathological type(x2=6.622,P ＜ 0.05),but not associated with infiltration depth(x2=4.914,P＞0.05).The expression of iNOS was associated with tumor size(x2 =8.609,P＜0.05),but was not associated withpathological type(x2 =4.789,P＞0.05)and infiltration depth(x2 =4.309,P＞0.05).The expression of COX-2 was associated with tumor size(x2 =7.320,P＜0.05),but was not associated with pathological type(x2 =2.772,P＞0.05)and infiltration depth(x2 =2.103,P＞0.05).The expression of HIF-1 α,iNOS,and COX-2 were significantly correlated with the expression of VEGF(r =0.943,1.000,0.986,P ＜ 0.05).The expression of COX-2 was significantly correlated with the expression of iNOS(r =0.986,P＜0.05).The expression of HIF-1 α was significantly associated with the expression of COX-2(r=0.986,P＜O.05).The expression of HIF-1α was significantly associated with the expression of iNOS(r=0.943,P＜0.05).Conclusions The expressions of the HIF-1 α,iNOS,COX-2 and VEGF protein are up-regulated in choroidal melanoma.They may play important roles in the angiogenesis of the choroidal melanoma.Assessment of the expression of HIF-1α,iNOS and COX-2 may be useful for the diagnosis and therapy of choroidal melanoma.