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口腔底部鳞状细胞癌中基质金属蛋白酶-2、基质金属蛋白酶-9及CD31/微血管密度的免疫组织化学评估

Immunohistochemical evaluation of MMP-2, MMP-9 and CD31/microvascular density in squamous cell carcinomas of the floor of the mouth.

作者信息

Monteiro-Amado Flávio, Castro-Silva Igor Iuco, Lima Cristina Jardelino de, Soares Fernando Augusto, Kowalski Luiz Paulo, Granjeiro José Mauro

机构信息

Hospital for Rehabilitation of Craniofacial Anomalies, University of São Paulo, Bauru, SP, Brazil.

出版信息

Braz Dent J. 2013;24(1):3-9. doi: 10.1590/0103-6440201301941.

DOI:10.1590/0103-6440201301941
PMID:23657405
Abstract

The aim of this study was to evaluate the immunoexpression of MMP-2, MMP-9 and CD31/microvascular density in squamous cell carcinomas of the floor of the mouth and to correlate the results with demographic, survival, clinical (TNM staging) and histopathological variables (tumor grade, perineural invasion, embolization and bone invasion). Data from medical records and diagnoses of 41 patients were reviewed. Histological sections were subjected to immunostaining using primary antibodies for human MMP-2, MMP-9 and CD31 and streptavidin-biotin-immunoperoxidase system. Histomorphometric analyses quantified positivity for MMPs (20 fields per slide, 100 points grade, ×200) and for CD31 (microvessels <50 µm in the area of the highest vascularization, 5 fields per slide, 100 points grade, ×400). Statistical design was composed by non-parametric Mann-Whitney U test (investigating the association between numerical variables and immunostainings), chi-square frequency test (in contingency tables), Fisher's exact test (when at least one expected frequency was less than 5 in 2×2 tables), Kaplan-Meier method (estimated probabilities of overall survival) and Iogrank test (comparison of survival curves), all with a significance level of 5%. There was a statistically significant correlation between immunostaining for MMP-2 and lymph node metastasis. Factors associated negatively with survival were N stage, histopathological grade, perineural invasion and immunostaining for MMP-9. There was no significant association between immunoexpression of CD31 and the other variables. The intensity of immunostaining for MMP-2 can be indicative of metastasis in lymph nodes and for MMP-9 of a lower probability of survival.

摘要

本研究旨在评估口腔底部鳞状细胞癌中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)和CD31/微血管密度的免疫表达情况,并将结果与人口统计学、生存率、临床(TNM分期)和组织病理学变量(肿瘤分级、神经周围浸润、栓塞和骨浸润)进行关联分析。回顾了41例患者的病历和诊断数据。组织学切片采用抗人MMP-2、MMP-9和CD31的一抗及链霉亲和素-生物素免疫过氧化物酶系统进行免疫染色。组织形态计量分析对MMPs阳性率(每张切片20个视野,100分制,×200)和CD31阳性率(血管化程度最高区域内直径<50 µm的微血管,每张切片5个视野,100分制,×400)进行量化。统计设计包括非参数曼-惠特尼U检验(研究数值变量与免疫染色之间的关联)、卡方频率检验(列联表分析)、费舍尔精确检验(当2×2表中至少一个期望频数小于5时)、Kaplan-Meier法(估计总生存率概率)和对数秩检验(比较生存曲线),所有检验的显著性水平均为5%。MMP-2免疫染色与淋巴结转移之间存在统计学显著相关性。与生存率呈负相关的因素包括N分期、组织病理学分级、神经周围浸润和MMP-9免疫染色。CD31的免疫表达与其他变量之间无显著关联。MMP-2免疫染色强度可提示淋巴结转移,而MMP-9免疫染色强度则提示生存率较低。

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