| ObjectiveLaryngeal carcinoma is a common malignant tumor in otorhinolaryngology, and the morbidity increases year by year. Designing a clinical scheme of treatment and selecting surgery approach lies on the size and extension of tumor. So it is important to make clear whether tumor is still localized in primary site or had metastasized to cervical lymph nodes or distant organs. At present, the methods used to evaluate the degree of invasion and metastasis of laryngeal carcinoma are physical examination, radiophotograph method and conventional pathological method, but they are not all accurate ways. It is necessary to find new tumor biomarkers to estimate it exactly. The process of tumor is very complicated, and it involves interaction of many factors at the level of cell and molecule. According to the Liofta's theory about metastasis of tumor, the process may comprise three steps: adhesion, degradation and migration. We know that it is the key step of breaking through the basement membrane (BM) during the invasive process. During the process of tumor diffusion, cancer cell can secrete itself and/or induce surrounding matrix cells to secrete type IV collagenase (including matrix metalloproteinase 2 and 9, or MMP-2 and MMP-9) to degrade type IV collagen in BM. Then tumor cells emigrate to the connective tissue and even distant organs. It has been proved that the expression level of type IV collagen decrease when the tumors become deteriorated such as breast cancer and bladder cancer. This phenomena is due to the unbalance between the secretion and the degradation of type IV collagen. Regulation of type IV collagenase involves several factors and works at three levels: (1) transcription; (2) activation of proenzyme; (3)post-activation. As a special inhibitor of type IV collagenase, tissue inhibitor of metalloproteinases(TIMPs) plays its role at (2) and (3) level. Few studies had been done about it in laryngeal carcinoma tissue. We made the experiment to ensure the relationship between the development, prognosis of laryngeal carcinoma and expression of type IV collagen, collagenase and TIMP-1.Materials and Methods1 Material1.1 Specimens of laryngeal carcinoma44 specimens were selected in this study that were collected from 1996 to 2000 in Hangzhou First Peoples Hospital. 41 cases were male, 3 female. They were from 46 to 84 years old, and the median age was 67. According to the TNM classification of UICC in 1997, there were 22 supraglottic cancers, 21 glottic ones and 1 subglottic one; Tl, T2, T3 and T4 tumors were 7, 9, 9 and 19 respectively; NO, Nl, N2 and N3 were 26, 10, 6 and 1 respectively. Well-, middle- and poor-differentiation were 10, 33 and 1 respectively. All specimens were diagnosed as laryngeal squamous cell carcinoma by HE stain.1.2 Specimens of para-tumor normal tissue22 cases of para-tumor were from samples of laryngeal carcinoma that were obtained from 1.0cm to 1.5cm to the border of tumor nest and were determined as normal tissue by HE stain. 20 cases were male and 2 female.2 Method2.1 I IE stainHE stain is used to confirm the character of tissue and the differentiation degree of laryngeal squamous cell carcinoma.2.2 Immunohistochemical methodImmuneohistochemical technique SP method was performed in ourexperiment to measure the expression of type IV collagen, MMP-2, MMP-9 and TIMP-1 in all specimens. All primary antibodies were monoclone antibodies (the concentration of first antibody of type collagen, MMP-2, MMP-9 and TIMP-1 was 1:100, 1:50,1:50 and 1:50 respectively). Positive control of type IV collagen was type IV collagen of BM of vessel endothelium or nerve membrane in the same slice, MMP-2, MMP-9 and TIMP-1 were known as positive slice of cancer which were obtained from Zymed Corporation. Negative control was obtained by replaced first antibody with PBS.2.2.1 Detection of stain resultType ]V collagen: 5 high views( 200) were detected in every section by two independent pathologists respectively. The positive parts local... |
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