The Observation Of Intratumoral And Peritumoral Lymphangiogenesis Vessel Density Of Cervical Lymph Node Metastasis In Nude Mice Bearing Human Tongue Squamous Cell Carcinoma

Object:1. This study using immunohistochemical method to mark the lymphatic of tongue squamous carcinoma lesion and lymph node metastasis,which to analysis the characteristics of tongue squamous cell carcinoma lymph angiogenesis and its correlation with clinicopathological index. Using lymphatic specific marker LYVE-1to mark the lymphatic of tongue squamous carcinoma, to calculation of lymphatic vessel density value,to study the correlation between lymphatic vessel density(LVD) of intratumoral and peritumoral and lymph node metastasis, along with the possible mechanism.2. To establish an orthotropic model of human tongue squamous cell carcinoma in nude mouse, simulate human body environment, observe the tumors cell biological behavior and condition of lymph node metastasis in vivo.Method:1. Detection of38cases by immuohistochemical method (male21case, female17cases) of lesion of tongue squamous cell carcinoma metastasis positive patients and24cases (including malel2case,femalel2case) expression of neck lymph node metastasis negative patients with the organization of the LYVE-1,using rabbit anti-human hyaluronidase-1(LYVE-1) marker to mark lymphatic,and count the lymphangiogenesis density of intratumoral and peritumoral.To analysis the characteristics of lymphtic distribution,and to study the relationship of lymphangiogenesis and clinicopathological index.Data is analyzed with SPSS13.0software package and T test is used for comparative analysis of intratumoral and peritumoral between lymph node metastasis and without lymph node metastasis.2. The Tca83cell were injected into the left margin of tongue in nude mouse. Every location was injected with105/0.05ml. To Check and record the weight, the condition of tumor growth, the length and width of tumor every two days. Mouse were sacrified when they died or their weight were less than25%of primay data. The tongue, lymph nodes were stained with hematoxylin and eosin for microscopic observation, and evaluated for histopathology.Result:1. The intratumoral exist isolated、neonatal、incomplete、tubulous structure, not with the surrounding host lymphatic endothelial eel gap is communicated,the tubulous structue can be seen nore,luminal narrow;the peritumoral which ones are larger and dilated.The peritumoral lymphatic vessels density with lymph node metastasis(14.500±3.122) was significantly higher than that without lymph node metastasis(9.083±3.744, P<0.05).By χ2test, the age, gender, location, size of the tumor have no significant difference between the lymph node metastasis and without lymph node metastasis(P>0.05); the intratumoral lymphatic vessels density with lymph node metastasis (7.500±2.784) was not significantly higher than that without lymph node metastasis(6.420±0.520, P>0.05).By T test, the lymphatic vessel density between age, gender, location,TNM, lymph node metastasis and without lymph node metastasis are significantly different.The peritumoral lymphatic vessel density of tongue squamous cell carcinoma with lymph node metastasis is signficantly related with lymph node metastasis and diatant organ metastasis.2. Tumor was formed in6th-11th day after injected tumor cell. Tumor formation rate was66.67%, the result of HE and Immunohisochemical showed cervical metastasis rate was36.11%. Both intratumoral and peritumoral have positive expression, showed lymphatic irregular shape, thin wall, can be in open state, cavity does not contain red blood cells, and blood vessels are obviously different, sometimes with jelly lymph; also can behave for lumen collapse, atresia were cord-like, even a single endothelial cells; sometimes visible lumen is not complete. The lymphatic vessels morphology of intratumoral and peritumoral without obvious difference, for the expanding shape. The peritumoral lymphatic vessels density with lymph node metastasis(5.943±2.020) was significantly higher than that without lymph node metastasis(3.875±1.260, P<0.05)。 Conclusion:1.(1) hyaluronic acid enzyme-1(LYVE-1) in tongue squamous cell carcinoma tissues and cervical lymph node metastasis,suggesting hyaluronic acid enzyme-1(LYVE-1) play an important role in the pathogenesis of tongue squamous cell carcinoma.(2) a large number of new lymphatic vessels in tongue squamous cell carcinoma.many distributed in the peritumoral.(3) The peritumoral lymphatic vessel density with lymph node metastasis is significantly higher than that without lymph node metastasis.(4) The peritumoral lymphatic vessel density of tongue squamous cell carcinoma with lymph node metastasis is signficantly related with lymph node metastasis and diatant organ metastasis. The lymphangiogenesis of tongue squamous cell carcinoma has any value to calculate lymph node metastasis.2. The establishment of a orthotropic mode human tongue squamous cell carcinoma in nude mouse was successful. It supplied an important experiment medium in studying tongue cancer development. Although injected Tca83cell in nude mouse didn’t have a ideal rate of lymph node metastasis, the result was more close to clinic characteristic and imitated the process of development of human head and neck carcinoma. The study provided useful information in establishing tongue cancer with high rate of lymph node metastasis.