| Tumor cell invasion and subsequent distant spread are the main characteristics of malignant tumors and the leading causes of their poor prognosis. There are three critical events required for malignant tumor invasion which is the initial step of tumor metastasis-cell adhesion, degradation of surrounding matrix and cell movement. Degradation is the most important factor during transformation from tumor in situ or pre-invasive tumor to invasive tumor.The main components of basement membrane(BM) are collegen IV, hepa-rin-like heparin sulfate protoglycans ( HSPG) , laminin and intracellular stroma. Tumor invasion was promoted by secretion of hydrolytic enzymes and degradation of tissue barrier. It is mentioned that BM degradation has a significant relationship with tumor invasion and metastasis, and its structure and distribution correlated with lymphatic metastasis. It is in need of secretion of hydrolytiv enzymes for squamous carcinoma of head and neck to break down barriers and possess invasion.Invasion and metastasis are continuous processs and need many factors in different stage. Interaction between tumor cell and parental cell may play the key role. It is mentioned that the production of molecular and enzymes on cell surface determined the invasion of cancer cells of head and neck squaumous carcinomas. Cytokines which conclude MMP-9, c-Met, heparanase ( hpa ) and VEGF, act as signals of proliferation, differentiation, activation and movement, communicating cellular interaction.Many researches have shown that hpa negatively expressed in normal tissues while expressed significant higher in many tumor tissues, which might be important for early diagnosis and prediction of patients prognosis. Very few reports were about the expression of hpa in human laryngeal carcinomas. It has been mentioned that abnormal transduction of receptors of HGF { or scatter factor,SF) had relationship with tumor proliferation, invasion and metastasis.PCNA, a nuclear protein emerged in cell nucleus from satge Gl to M, is a factor for cell proliferation. It may reflect the character of cell growth and had an important role in prognosis. MMPs, a family of ECM degradation enzymes, is significantly related with angiogenesis and tumor metastasis.In the current experiment, we detected pathological factors in 54 laryngeal carcinomas and 10 adjacent normal tissues by use of immunohistochemistry and in situ hybridizaition to evaluate the significance of hpa, c-Met, PCNA and MMP-9 with clinical pathological factors and tumor invasion and metastasis. We also detected the micrometastasis of reactive hyperplasia lymph nodes and analyse its role in clinical diagnosis.Materials and methodsTissue SamplesFifty-four human laryngeal carcinoma (HLCC) tissue samples and 31 reactive hyperplasia lymph nodes included in the current study were obtained from 34 male and 20 female patients (median age, 58 years; range, 48-78years) underwent surgical resection in the first affiliated hospital of china medical university between 1996 and 1997. Together with the cancer tissues, adjacent normal lung tissues in 10 patients were resected. No patient in this study had been pr-scribed chemotherapy or radiotherapy prior to surgery.ImmunohistochemistryFormalin-fixed and paraffin-embedded tissue sections (4um thick) were deparaffinized and rehydrated, antigen retrieval was performed by citrate buffer (pH 6). Sections were incubated with the heparanase , PCNA, P-CK and MMP-9 antibodies, respectively. Immunohistochemical staining was performedusing a kit according to the instructions provided by the manufacturer. Color was de veloped using DAB/H2O2 solution. Specimens were incubated with PBS instead of primary antibody as control.In Situ HybridizationParaffin-embedded specimens (4um) were deparaffinized and rehydrated. The steps of hybridization were performed conducted using a kit according to the recommendations of the manufacturer.Results1. Expression of hpa expression. Of 54 cancer tissues used for immunohis-tochem...
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