胃癌CD^＋4 CD^high 25 Foxp3^＋调节性T淋巴细胞数量和分布及Foxp3基因表达与肿瘤分期的相关性  

The populations, distribution of regulatory T cells, Foxp3 mRNA expression gastric cancer patients and the association with malignant stage

作　　者：袁向亮[1,2] 沈定丰[3] 卢剑[1,2] 董平[3] 王剑[1,2] 李美星[3] 沈立松[1,2] 

机构地区：[1]上海交通大学医学院附属新华医院,200092 [2]上海儿童医学中心临床实验诊断中心,200092 [3]上海交通大学医学院附属新华医院普外科,200092

出　　处：《中华检验医学杂志》2008年第4期378-383,共6页Chinese Journal of Laboratory Medicine

基　　金：基金项目：上海教育委员会自然科学基金资助项目（06B2050）

摘　　要：目的 研究胃癌患者CD^＋4 CD^high 25 Foxp3^＋调节性T淋巴细胞（Treg）的存在数量、分布情况，及其功能基因Foxp3的表达变化与胃癌临床国际肿瘤淋巴转移分类（TNM）分期的关系。方法 用四色流式细胞术以Foxp3·FITC／CD25-PE／CD4-PerCP／CD3-PC7抗体组合分析20名健康对照者及47例胃癌患者（临床TNM分期Ⅰ11例、Ⅱ期18例、Ⅲ期10例、Ⅳ期8例）外周血、腹腔积液、肿瘤组织浸润淋巴细胞（TIL）及癌旁淋巴结中CD^＋4 CD^high 25 Foxp3^＋ Treg的数量及分布情况。实时荧光定量PCR技术检测Treg Foxp3 mRNA表达水平。结果 胃癌患者外周血CD^＋4 CD^high 25 Foxp3^＋ Treg占CD^＋4 T淋巴细胞的百分比为（8．03±3．47）％，高于正常健康对照组（5．83±2．93）％，差异具有统计学意义（P〈0．05）。胃癌患者腹腔积液、肿瘤浸润淋巴细胞、癌旁淋巴结中Treg占CD^＋4 T淋巴细胞比例均高于外周血，差异有统计学意义（分别为P〈0．05、P〈0．01、P〈0．01）。胃癌组织局部，TIL和癌旁淋巴结中Treg的Foxp3平均荧光强度显著高于外周血和腹腔积液（P〈0．05）。Ⅰ、Ⅱ、Ⅲ、Ⅳ期胃癌患者Treg比例分别为（5．72±1．95）％、（6．45±2．23）％、（9．58±3．13）％、（11．70±2．30）％，这提示Treg与肿瘤TNM分期显著相关（r=0．784，P〈0．01）。胃癌患者外周血单个核细胞Foxp3基因mRNA表达水平明显高于健康对照组（P〈0．01）。Foxp3基因表达水平与胃癌TNM各分期呈显著相关性（r=0．623，P〈0．05）。结论 胃癌患者CD^＋4 CD^high 25 Foxp3^＋ Treg的比例升高，其增高程度与肿瘤临床分期呈显著正相关。胃癌患者外周血和癌组织局部Treg的数量、分布情况及特异性转录因子Foxp3的表达强度均有所不同，提示肿瘤局部免疫抑制功能增强，这可能是产生肿瘤免疫抑制的重要机制，CD^＋4 CD^high 25 Foxp3^＋Treg将来可作为胃癌�Objective To investigate the populations of CD^＋4 CD^high 25 Foxp3^＋ regulatory T cells （Treg cells） and mRNA expression of Foxp3 in peripheral and the marginal region of tumor from patients with gastric cancer, and to evaluate the prevalence of Treg cells from patients with gastric cancer in relation to the TNM stages. Methods PBMC, Ascites, tumor-infiltration lymphocyte and tumor-draining lymph nodes in 47 patients with gastric cancer （TNM Ⅰ 11, Ⅱ18, Ⅲ10, Ⅳ 8） and PBMC in 20 normal healthy donors were evaluated for the proportion of Treg cells, as a percentage of the total CD^＋4 cells, by flow cytometric analysis. Levels of mRNA for Foxp3 were measured with a real-time quantitative PCR. Results The percentage of CD^＋4 CD^high 25 Foxp3^＋ Treg cells in PBMC for cases of gastric cancer were significantly higher than those for healthy donors （ P 〈 0. 05 ）. The percentage of Treg cells were more abundant in ascites （ P 〈 0. 05 ）, TIL （ P 〈 0. 01 ） and tumor-draining lymph nodes （ P 〈 0. 01 ） of individuals with gastric cancer than in their blood. And the Foxp3 mean fluorescent intensity （MFI） of Treg cells in TIL and tumor draining lymph nodes with gastric caners were significantly higher than PBMC and ascites （P 〈 0. 05 ）. The proportion of Treg cell in patients of gastric cancers with stage Ⅰ , Ⅱ ,Ⅲ, Ⅳ were （5.72 ± 1.95 ） %, （6.45 ± 2. 23 ） %, （9. 58 ± 3. 13 ） %, （ 11.70 ± 2. 30 ） %, respectively. There were significant correlation between prevalence of Treg cells and disease stages in gastric cancer （ r = 0. 784, P 〈 0. 01 ）. Foxp3 mRNA levels were higher in PBMC from the group with gastric cancers than in the group with healthy donors. Foxp3 mRNA levels were correlated with TNM stages （ r = 0. 623, P 〈 0. 05 ）. Conclusions Our results suggest that the populations of CD^＋4 CD^high 25 Foxp3^＋ Foxp3 Treg cells and Foxp3 mRNA levels in patients with gastric cancer are significantly higher in comparison to those i

关 键 词：胃肿瘤 肿瘤分期 DNA结合蛋白质类 T淋巴细胞 

分 类 号：R686[医药卫生—骨科学]