Preliminary Study On The Roles Of Chemokine Receptor CCR7and Aquaporin-1in Metastasis Of Human Tongue Carcinoma

Metastasis,which may occur either by the blood vascular system or by dissemination via the lymphatic vasculature, is a completion of a multi-process characterized by highly organized, non-random and organ-specific. The molecular mechanism of the directional migratory of tumor cells towards lymph nodes continue to be elucidated although varies of cytokines involved in the progress of tumor invasion and migration are revealed. To date, studies showed the interaction between chemokine receptor CCR7and ligands may plays a crutial role in the tumor cells migration to lymph nodes whereas there were few reports described the roles both CCR7and its ligands in human tongue carcinoma, much less of the mechanism. To investigate the the role of CCR7/CCL21in metastasis to lymoh nodes in tongue carcinoma, we delected the expression of CCR7/CCL21in both tongue carcinom and metastatic lymph nodes using IHC and evaluated the correlation with clinicopathologic characteristics and prognosis. Moreover, a zebrafish tumor xenograft assays was established to observe the clinical situation of metastasis in both CCR7-silenced zebrafish by RNAi technic and negative control which supplied the basis to further investigation of the role ofCCR7in tongue carcinoma metastasis.Angiogenesis is not only essential for primary tumor growth but also facilitates tumor infiltration and metastasis. A developing body of evidence showed hypoxia in tissues and cells plays a critical role in the development of angiogenesis. It is widely studied that the aquaporins family and hypoxia-induced factors involve in the process of tumor metastasis caused by hypoxia. Of which, the correlation between AQP1with HIF-1α expression in human tongue carcinoma and metastatic lymph nodes is rarely reported. In this study, we performed IHC to detective the expression level of both AQP1and HIF-1α and proposed a possibility of the relations with clinicopathotogic factors which supplied the basis to further investigation of the roles AQP1pkys in human tongue carcinomaPart One Correlation of CCR7expression with metastasis and prognosis of tongue carcinomaObjective:To investigate the expression of CCR7in the tongue cancer and analyse the correlations between CCR7expression and clinicopatho logical factors of tongue cancer metastasis and prognosis. Methods:we investigated the role of CCR7in the lymph node metastasis of tongue carcinoma in current study. Immunohistochemistry and Western blot revealed the expression of CCR7in tongue SCC tissues and cell lines. Moreover, we detected the expression level of CCL21, a ligand of CCR7, in normal and diseased lymph nodes using immunohistochemistry and/or real-time PCR. Results:The CCR7expression was significantly correlated with cervical lymph node metastasis, tumor staging, and histological grade (P=0.015,0.040, and0.015, respectively). The multivariate analysis showed that regional lymph node metastasis, the expression of CCR7, and LVD were the independent poor prognostic factors. CCR7knockdown obviously inhibited tumor growth, dissemination and metastasis in an tumor xenograft zebrafish model. Conclusion: Our study indicated that CCR7may play an important role in progression of tongue SCC and could be a promising target for tongue SCC therapy. Part Two Role of aquaporin1expression in metastasis and tumor angiogenesis of tongue carcinomaObjective:To determine the expressions of aquaporin1(AQP1) and Hypoxia-inducible factor-la (HIFla) in relation to angiogenesis (micro vessel density, MVD) and clinicopathologic factors in human tongue carcinoma. Methods: Immunohistochemical staining was used to detect the protein expression of AQP1and HIF1α in paraffin-embbeded tissue samples surgically resected from74tongue cancer patients and10normal mucosa tissues. Results:In all cases of human tongue carcinoma, MVD was evaluated by counting CD34-reactive endothelial cells or endothelial cell clusters.AQPl is expressed on the membrane of microvessels and small vessels in all samples, with a mean score of20.73±7.22(mean±SD), with showing in neither normal mucosa epithelium nor tongue carcinoma cells. Of the74human tongue carcinoma samples,30showed mainly nuclear, but also cytoplasmic HIF1α expression. AQP1expression had significant correlations with MVD and HIF1α expression (P=0.000and0.002, respectively). The AQP1, HIF1α and MVD were significantly correlated with clinical stage (P=0.000,0.012and0.000, respectively) and lymph node metastasis (P=0.000,0.011and0.000, respectively). Conclusion:The overexpressions of AQP1and HIFla were involved in both angiogenesis and progression of human tongue carcinoma. This study suggests that the expression of AQP1and HIF1α may be important biomarkers prognosis for human tongue carcinoma.