| Background and ObjectiveGrowth and metastasis of solid tumors depend on angiogenesis. The presence of an angiogenic process implies the existence of regulators of the process. Many angiogenic cytokines have been proven to be involved in this process, serving as the positive regulators. Among them, Vascular endothelial growth factor (VEGF) is considered to be the most critical one, which can exert different functions in every step of angiogenesis. Matrix metalloproteinases(MMPs) are a family of zinc-dependent endopeptidases collectively capable of degrading extracellular matrix(ECM) during angiogenesis, invasion and metastasis of tumor. Numerous studies have shown that different cytokines always promote angiogenesis cooperatively, so it is necessary to investigate several cytokines in the same tumor tissue for understanding the whole process of angiogenesis. At present, therapeutic intervention of tumor angiogenesis based on inhibition of VEGF/MMP activity is also under intensive investigation, moreover, some inhibitors have met with remarkable success in clinical trials. Laryngeal cancer is one of the most common carcinomas of the head and neck, being caused by both bodily and environmental carcinogenic factors. Angiogenesis might be related to the malignant progression of laryngeal cancer, but there have been few investigations in this field. Here the expressions of VEGF, MMP-9 and microvesselcount (MVC) were detected by immunohistochemical method in 42 laryngeal carcinoma samples. The purpose of this study is to explore the relationship among VEGF or MMP-9 expression and the angiogenesis or clinicopathological characteristics of laryngeal cancer. It may provide data for evaluating the prognosis of laryngeal cancer, as well as the potential antiangiogenic therapy for laryngeal cancer.Materials and Methods42 samples were obtained from surgical specimens of the patients with laryngeal cancer hospitalized in the First Affiliated Hospital, Medical College, Zhejiang University during the period 1997-2002.Among these patients, 39 were male and 3 were female. The average age was 57.6±7.79 years. According to the TNM classification of UICC in 1997, 42 patients were separated into I , II ,III and IV stage, 9, 11, 18 and 4 respectively. Histologically, tumors were graded as well, moderately and poorly differentiated, 20, 17 and 5 respectively. All these patients had not received preoperative chemotherapy or radiotherapy. In addition, all these samples were diagnosed to be laryngeal squamous cell carcinoma (LSCC) by HE dying.VEGF, MMP-9 and CD34 expression were detected by immunohistological method in all the samples. The results were observed under light microscope. The degree of VEGF/MMP-9 expression was graded as negative or slightly-positive expression (stained tumor cells/all counted tumor cells<25%) and positive expression (>25%). MVC was obtained by counting the number of CD34-positive microvessel or isolated endothelial cells and clusters of endothelial cells.The data were analyzed by statistical package (SPSS 10.0 for windows). The statistical methods include t-test, chi-square test and Fisher's exact test.Results1. VEGF positive tumors showed a significant higher mean MVC than VEGF negative or slightly-positive tumors (23.85±3.70/19.63±5.45,P=0.012). The expression of VEGF was not correlated with the differentiation and TNM stage of LSCC2. MMP-9 positive tumors showed a significant higher mean MVC than MMP-9 negative or slightly-positive tumors (24. 19±4.09/21.00±4.17,P=0.021). The expression of MMP-9 was not correlated with the differentiation and TNM stage of LSCC (P>0.05).3. The cases of VEGF(+)/MMP-9(+),VEGF(+)/MMP-9(-),VEGF(-)/MMP-9(+) and VEGF(-)/MMP-9(-) expression pattern were 23, 11, 4 and 4 respectively. VEGF expression and MMP-9 expression showed no significant correlation in LSCC (P=0.425).Conclusions1. VEGF and MMP-9 expression are significantly associated with MVC in LSCC. They may play an important role in the angiogenesis of...
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