CCR6、CCR7及CD4(+)CD25(+)Foxp3(+)调节性T细胞在喉鳞状细胞癌及颈部淋巴结转移中的表达及临床意义

Chen Bin, Lu Li-ming, Tao Lei, Zhou Liang, Li Shi-min, Zhu Li

Department of Otolaryngology, Eye Ear Nose & Throat Hospital of Fudan University, Shanghai 200031, China.

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2010 Sep;45(9):759-64.

OBJECTIVE

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 and fresh specimens of LSCC were obtained from 50 LSCC patients treated. Blood samples from normal subjects were donated by 20 volunteers. The mRNA expressions of CCR6, CCR7 and their ligands CCL20, CCL19/CCL21 were detected by real-time quantitative RT-PCR. Expressions of CCR6 and CCR7 proteins were examined with immunohistochemistry. Flow cytometry was used to investigate Treg in peripheral blood mononuclear cells.

RESULTS

The relative expression levels of CCR6, CCR7 and CCL19 mRNA in tumor tissues with lymphatic metastases were significantly lower than those without lymph node metastases (P < 0.05), while the relative expression level of CCL20 mRNA in tumor tissues with lymphatic metastases was significantly higher than that of those without metastatic lymph nodes (t = 2.39, P < 0.05). Immunohistochemistry showed that the expression levels of CCR6 and CCR7 in tumor tissues in patients with lymph node metastases than those without lymph node metastases. CCR6 and CCR7 expressions were also detected in metastatic lymph nodes. Flow cytometry showed that the percentage of Treg in blood in LSCC patients was significantly higher than that in normal subjects (t = 2.19, P < 0.05), and those with lymph node metastasis had a much higher percentage of Treg (t = 2.14, P < 0.05).

CONCLUSIONS

CCR6, CCR7 and Treg could take part in the process of lymph node metastasis in LSCC patients.

目的

评估趋化因子受体6（CCR6）、趋化因子受体7（CCR7）及CD4(+)CD25(+)Foxp3(+)调节性T细胞（Treg）在喉鳞状细胞癌（LSCC）及转移淋巴结中的表达情况及其临床意义。

方法

收集50例接受治疗的LSCC患者的血液样本及新鲜肿瘤标本。20名志愿者捐献了正常受试者的血液样本。采用实时定量逆转录聚合酶链反应（RT-PCR）检测CCR6、CCR7及其配体CCL20、CCL19/CCL21的mRNA表达。采用免疫组织化学法检测CCR6和CCR7蛋白的表达。采用流式细胞术检测外周血单个核细胞中的Treg。

结果

有淋巴结转移的肿瘤组织中CCR6、CCR7及CCL19 mRNA的相对表达水平显著低于无淋巴结转移者（P < 0.05），而有淋巴结转移的肿瘤组织中CCL20 mRNA的相对表达水平显著高于无转移淋巴结者（t = 2.39，P < 0.05）。免疫组织化学显示，有淋巴结转移患者肿瘤组织中CCR6和CCR7的表达水平低于无淋巴结转移者。在转移淋巴结中也检测到了CCR6和CCR7的表达。流式细胞术显示，LSCC患者血液中Treg的百分比显著高于正常受试者（t = 2.19，P < 0.05），有淋巴结转移者Treg的百分比更高（t = 2.14，P < 0.05）。