| ObjectiveLaryngeal carcinoma is a common malignant tumor in Otolaryngology. Its morbidity was increased year by year. Early diagnosis and treatment help to improve patient's survivor probability. Some studies had proved that tumor growth and metastasis were angiogenesis dependent. Growth and metastatic dissemination of solid tumors require vascular support for nutritive supply and access. Apoptosis is a genetically encoded program of cell death that serves as a central regulation for the development and homeostasis of multicellar organisms. Tumor not only had abnormal proliferation but also had abnormal apoptosis. The inhibitors of angiogenesis can limit tumor growth and metastasis by elevating the incidence of apoptosis in tumor cells. Recently, some studies have demonstrated that angiogenesis had significantly relationships with tumor growth, progression, metastasis and prognosis. Intratumor microvessel density(IMVD) was a important indicator of tumor prognosis. However, study on the relationship among angiogenesis, apoptosis and clinicopathological features, prognosis in LSCC is rare; and data on this topic are still controversial. It is necessary to study further. In this experiment,we investigate expression of tumor angiogenesis, vascular endothelial growth factor(VEGF) by immunohistochemical methods and spontaneous apoptosis by TUNEL(terminal-deoxynucleotidyl transferase mediated nick end labeling) method in 34 patients with LSCC, to find out the relationship among angiogenesis, VEGF expression, spontaneous apoptosis and clinicopathological chacteristics. It may provide data for evaluating progression, prognosis, antiangiogenesis treatment in LSCC.Materials and MethodsIn this study, 34 patients who were treated for laryngeal cancer were from 1999 to 2000 in the first affiliated hospital of medical college, Zhejiang University. 30 patients were male, and 4 were female. Their age ranged from 48 to 77, and the average age was 59.4 + 6.2. According to the TNM classification of UICC in 1992, there were 7 supraglottic cancers, 26 glottic ones and 1 subglottic one. Tl tumors were 6, and T2, T3, T4 tumors were 13, 11,4 respectively. Among all 34 patients, there were 15 patients with cervical lymph node metastasis. All were proved to be squamous cell carcinoma by biopsy.In this experiment, we use immunohistochemical SABC(StreptAvidin-Biotin Complex) method to detect angiogenesis of LSCC tissues by CD34 monoclonal antibody and VEGF expression by VEGF polyclonal antibody, and TUNEL(terminal deoxynucleotidyl transferase mediated nick end labeling) method to detect spontaneous apoptosis of laryngeal cancer cells. The results were observed in light microscope. Average IMVD of each slide was counted by CD34 positive stain cell and VEGF expression by VEGF positive tumor cells/all counted tumor cells. Apoptosis index was counted by positive tumor cells/all counted tumor cells.The data were analyzed by statistical package(SPSS for windows). Main statisticalmethods were t-test, chi-square test, Fisher exact test, Spearman rank correlation and analysis of variance.Results1. AH 34 slides were positive stained by CD34. Average IMVD was 21.5 + 8.87. IMVD had a statistical significant relevance with cervical lymph node metastasis of LSCC patients (P<0.01); Tumors with cervical lymph node metastasis had a significantly higher IMVD than those without cervical lymph node metastasis. Tumors with average IMVD^21.5 had a significantly higher risk of cervical lymph node metastasis than those with IMVD<21.5 (P<0.05). IMVD revealed a reverse correlation with histological grading (P<0.01). No correlation was found between IMVD and tumor T stage(P>0.05).2. Average IMVD had a positive relationship between patients with VEGF higher expression and those with lower expression (P<0.05). Tumors with VEGF higher expression had no significantly higher risk of cervical lymph node metastasis than those with lower expression(P>0.05).3. Apoptosis index of laryngeal cancers ranged from 0% to 5.8%. Apoptosis ind...
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