软骨肉瘤中的一氧化氮合酶、环氧化酶-2、硝基酪氨酸和血管生成及其与预后的关系。

Nitric oxide synthases, cyclooxygenase-2, nitrotyrosine, and angiogenesis in chondrosarcoma and their relation to prognosis.

机构信息

Departamentos de Cirurgia Pélvica-Ortopedia, Hospital A.C. Camargo, Rua Prof. Antonio Prudente, 211, São Paulo - S. P., CEP 01509-010, Brazil.

出版信息

J Bone Joint Surg Am. 2010 Jul 21;92(8):1738-46. doi: 10.2106/JBJS.H.00717.

DOI:10.2106/JBJS.H.00717

PMID:20660237

Abstract

BACKGROUND

The localization in tumor tissue of various markers by immunohistochemistry can help to establish a diagnosis or predict prognosis. Nitric oxide is associated with tumors and has been studied indirectly by nitrotyrosine analysis and with use of the enzymes nitric oxide synthase (NOS)1, NOS2, and NOS3. Nitric oxide reacts with superoxide anions to yield peroxynitrite, which has toxic effects on genes. Peroxynitrite adds a nitro group to the benzene ring of tyrosine to form nitrotyrosine. The accumulation of nitrotyrosine, a stable product in cells, indicates the formation of peroxynitrite. Nitric oxide stimulates the production of cyclooxygenase-2 (COX-2), which has been associated with angiogenesis in tumors. Neovascularization influences tumor prognosis, as demonstrated by microvessel studies with use of CD34, an immunohistochemical endothelial cell marker. This study examines the expression of these markers in chondrosarcomas and their relation to histological grade and prognosis.

METHODS

Tissue microarrays composed of formalin-fixed tissue samples from 101 patients with chondrosarcoma were immunohistochemically stained to localize NOS1, NOS2, NOS3, COX-2, nitrotyrosine, and CD34. Five samples of normal cartilage were used as controls. Patient demographics, selected surgical variables, and tumor grade were tabulated, and the associations were analyzed. Analyses of local and overall survival rates were performed with use of the Kaplan-Meier method, and multivariable analyses were performed.

RESULTS

There was a significant association of nitrotyrosine, COX-2, and CD34 with histological grades (p = 0.022, p = 0.014, and p = 0.028, respectively), but not with overall prognosis (p = 0.064, p = 0.143, and p = 0.581, respectively). The presence of NOS2 was associated with a lower rate of local disease-free survival (p = 0.038), and positive expressions of NOS1 and NOS2 were associated with decreased overall survival rates (p = 0.007 and p < 0.001, respectively). On multivariable analysis, NOS2 expression demonstrated an independent prognostic impact on local disease-free survival; NOS1 and NOS2 expression was a dependent variable, and their isolated or combined expression was related to lower overall survival rates (p = 0.046 and p = 0.004) (hazard ratio, 3.17 [95% confidence interval, 1.0 to 9.8] and 5.58 [95% confidence interval, 1.7 to 18.0], respectively).

CONCLUSIONS

Immunohistochemical markers may have an independent value in predicting the prognosis for patients with chondrosarcoma.

摘要

背景

通过免疫组织化学技术在肿瘤组织中定位各种标志物有助于确立诊断或预测预后。一氧化氮与肿瘤有关，曾通过硝基酪氨酸分析和使用一氧化氮合酶（NOS）1、NOS2 和 NOS3 进行间接研究。一氧化氮与超氧阴离子反应生成过氧亚硝酸根，后者对基因有毒性作用。过氧亚硝酸根将硝基加到酪氨酸的苯环上形成硝基酪氨酸。硝基酪氨酸在细胞中是一种稳定的产物，其积累表明过氧亚硝酸根的形成。一氧化氮刺激环氧化酶-2（COX-2）的产生，COX-2 与肿瘤中的血管生成有关。新血管生成影响肿瘤的预后，这一点可以通过使用 CD34（一种免疫组织化学内皮细胞标志物）进行微血管研究得到证明。本研究检测了软骨肉瘤中这些标志物的表达及其与组织学分级和预后的关系。

方法

对来自 101 例软骨肉瘤患者的福尔马林固定组织样本的组织微阵列进行免疫组织化学染色，以定位 NOS1、NOS2、NOS3、COX-2、硝基酪氨酸和 CD34。使用 5 例正常软骨作为对照。记录患者人口统计学资料、选择的手术变量和肿瘤分级，并进行关联分析。使用 Kaplan-Meier 方法进行局部和总生存率的分析，并进行多变量分析。

结果

硝基酪氨酸、COX-2 和 CD34 与组织学分级显著相关（p = 0.022、p = 0.014 和 p = 0.028），但与总体预后无关（p = 0.064、p = 0.143 和 p = 0.581）。NOS2 的存在与局部无病生存率降低相关（p = 0.038），NOS1 和 NOS2 的阳性表达与总生存率降低相关（p = 0.007 和 p < 0.001）。多变量分析显示，NOS2 表达对局部无病生存率具有独立的预后影响；NOS1 和 NOS2 表达是一个因变量，它们的单独或联合表达与较低的总生存率相关（p = 0.046 和 p = 0.004）（危险比分别为 3.17（95%置信区间为 1.0 至 9.8）和 5.58（95%置信区间为 1.7 至 18.0））。