Expression And Significance Of Ccr6, Ccr7 And Cd4~+cd25~+foxp3~+ Regulatory T Cells In Laryngeal Squamous Cell Carcinoma And Its Progression

Part I Expression of CCR6 and CCR7 in laryngeal squamous cell carcinomaObjective:To evaluate the expression of chemokine receptor 6 (CCR6), chemokine receptor 7 (CCR7) and their ligands (CCL20, CCL19/CCL21) in laryngeal squmaous cell carcinoma (LSCC) and explore their correlation with the clinicopathological features of LSCC. Methods:Blood samples, fresh specimens of LSCC and paired adjacent tissues were collected. The expression of CCR6, CCR7 and their ligands CCL20, CCL19/CCL21 mRNA as well as the protein CCR6, CCR7 were detected by real-time qRT-PCR and IHC respectively. Flow cytometry was also used to investigate CCR6, CCR7 expressed on PBMC. Results:The relative expression levels of CCR6, CCR7, CCL19 and CCL21 mRNA in tumor tissue was significantly higher than that of adjacent tissues (P<0.05), while the relative expression level of CCL20 mRNA in tumor situ was significantly lower than that of adjacent tissues (P<0.05). IHC confirmed the expression of protein CCR6 and CCR7 in both tumor situ and metastatic LN and the expression level of protein CCR6, CCR7 was higher in the cases with lymphatic metastasis than that of those without lymphatic metastasis (P<0.05). FCM showed the percentage of CD4+CCR6+T cells of LSCC was significantly higher than that of normal control (P<0.05),while that of CD4+CCR7+T cells was significantly lower (P<0.05). Conclusion:CCR6, CCR7 can be expressed in tumor situ, metastatic LN and PBMC, which might suggesting an potential role in LSCC development. Part II Expression and clinical significance of CCR6, CCR7 and CD4+CD25+Foxp3+ regulatory T cells in laryngeal squamous cell carcinoma and its lymphatic metastasisObjective:To evaluate the expressions and clinical significance of chemokine receptor 6 (CCR6), chemokine receptor 7 (CCR7) and CD4+CD25+Foxp3+ regulatory T cells (Treg) in laryngeal squamous cell carcinoma (LSCC) and metastatic lymph nodes. Methods:blood samples, fresh specimens of LSCC were obtained from 50 LSCC patients treated in our hospotal and blood samples from normal subjects were donated by 20 volunteers in our hospotal. The expressions of CCR6, CCR7 and their ligands CCL20, CCL19/CCL21 mRNA as well as the protein CCR6, CCR7 were detected by real-time quantitative RT-PCR (real-time qRT-PCR) and immunohistochemistry (IHC) respectively. Flow cytometry (FCM) was used to investigate Treg in peripheral blood mononuclear cell (PBMC). Results:The relative expression levels of CCR6, CCR7, CCL19 mRNA of tumor tissue from cases with lymphatic metastasis were significantly lower than that of those without lymph node metastasis (P<0.05), while the relative expression level of CCL20 mRNA in tumor was significantly higher than that of those without metastatic lymph nodes (F<0.05). IHC showed that the expression levels of CCR6 and CCR7 in tumor tissue were significantly higher in patients with lymph nodes metastasis than that of cases without lymph nodes metastasis. And the CCR6, CCR7 expression were also detected in metastatic lymph nodes. Flow cytometry showed the percentage of Treg in LSCC was significantly higher than that of normal subjects (P<0.05) and those with lymph nodes metastasis had much higher percentage of Treg (P<0.05). Conclusion:It was speculated that the expression profile of CCR6, CCR7 and the proliferation of Treg would have taken part in the process of lymph nodes metastasis in LSCC patients.