趋化因子CCL5在食管癌组织中的表达及意义  

Expression and clinical significance of CCL5 in patients with esophageal carcinoma

作　　者：刘金燕[1] 李峰[1] 陈新峰[1] 王丽萍[2] 岳冬丽[1] 赵松[3] 胡伟[3] Pawel Kalinski Stephen Thorne Hou Jingzhou 张毅[1] 

机构地区：[1]郑州大学第一附属医院生物细胞治疗中心,450052 [2]郑州大学第一附属医院肿瘤科,450052 [3]郑州大学第一附属医院胸外科,450052 [4]美国匹兹堡大学癌症研究中心 [5]美国匹兹堡大学癌症医学中心

出　　处：《中华肿瘤杂志》2014年第11期828-833,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金中美生物医学合作研究项目（81261120402）;国家自然科学基金（81171986）;卫生部科研攻关基金（2011010001）

摘　　要：目的 探讨趋化因子CCL5在食管癌组织中的表达及意义.方法 采用逆转录聚合酶链反应（RT-PCR）检测食管癌组织和癌旁组织中CCL5、CD8和CD8^＋T细胞杀伤功能相关细胞因子穿孔素（perforin）和颗粒酶B（granzyme B）的表达水平.采用流式细胞术检测食管癌患者外周血单核细胞（PBMC）和肿瘤浸润性淋巴细胞（TIL）中CD8^＋T细胞和CCR5^＋CD8^＋T细胞比例.采用Transwell实验检测CCL5对T细胞运动的影响.结果 食管癌组织中CCL5和perforin mRNA的表达水平分别为0.348 2±0.300 1和0.181 9±0.118 6,癌旁组织中CCL5和perforin mRNA的表达水平分别为0.279 6±0.138 0和0.118 0±0.1098,但差异均无统计学意义（均P＞0.05）.食管癌组织中CD8和granzyme B mRNA的表达水平分别为0.4649±0.3008和0.6487±0.5160,癌旁组织中CD8和granzyme B mRNA的表达水平分别为0.279 0±0.173 4和0.469 7±0.259 1,差异有统计学意义（均P ＜0.05）.食管癌患者中CCL5与CD8、perforin和granzyme B表达呈正相关（rCD8=0.272,P=0.034;rerforin=0.305,P=0.026;rgranzymeB=0.108,P=0.012）.流式细胞术结果显示,TIL和PBMC中CD8^＋T细胞比例分别为（45.86±16.09）％和（34.05±15.07）％,差异有统计学意义（P =0.022）;TIL和PBMC中CCR5^＋ CD8^＋T细胞比例分别为（48.12±26.75）％和（19.53±13.67）％,差异有统计学意义（P＜0.001）.Transwell实验结果显示,CCL5显著增强T细胞趋化运动.CCL5表达与患者性别、年龄和淋巴结转移无关,但CCL5在早期食管癌组织中的相对表达水平为0.319 9±0.161 7,在晚期食管癌组织中的相对表达水平为0.232 8±0.121 0,差异有统计学意义（P=0.008）.早期食管癌患者TIL中CD8^＋T细胞和CCR5^＋ CD8^＋T细胞比例分别为（48.86±15.87）％和（56.23±26.23）％,而在晚期食管癌患者TIL中CD8^＋T细胞和CCR5^＋ CD8^＋T细胞比例分别为（33.25±16.49）％和（33.53±21.03）％,差异均有统计学意义（PCD8 =0.007,PccR5=0.010）.结论Objective To investigate the expression and significance of CCL5 in patients with esophageal carcinoma.Methods Using reverse transcriptase polymerase chain reaction （RT-PCR）,the expressions of CCL5/CD8/granzyme B/perforin in tumor and corresponding adjacent tissues from esophageal carcinoma patients were examined.Flow cytometry （FACS） was used to detect the percentages of CD8 ＋ T cells and CCR5 ＋ CD8 ＋ T cells in TIL and PBMC from the patients.Transwell assay was performed to study the effect of CCL5 on the migration of T cells in vitro.T test and Spearman correlation analysis were performed.Results The mRNA expressions of CCL5 and perforin were 0.348 2 ±0.300 1 and 0.181 9 ± 0.118 6,respectively,in the tumor samples,while their expressions in adjacent samples were 0.279 6 ± 0.138 0 and 0.118 0 ± 0.109 8,respectively,with no statistically significant differences between them （P 〉 0.05 for both）.The mRNA expressions of CD8 and granzyme B were significantly higher in the tumor tissues than in adjacent tissues （0.464 9 ± 0.300 8 vs.0.279 0 ± 0.173 4,0.648 7 ± 0.516 0 vs.0.469 7 ± 0.259 1 ; P 〈 0.05 for both）.The relative expression of CCL5 was positively correlated with that of CD8,perforin and granzyme B （rCD8 =0.272,P =0.034; rperforin =0.305,P =0.026; rgranzymeB =0.108,P=0.012） in the tumor sites.FACS data revealed that the proportions of CD8 ＋ T cells in TIL and PBMC were （45.86 ± 16.09） % and （34.05 ± 15.07） %,respectively,showing a significant difference （P =0.022）.Similarly,CCR5 ＋ CD8^＋ T cells fraction in TIL （48.12 ± 26.75）% was much higher than that in PBMC（19.53 ± 13.67） % （P 〈 0.001）.Transwell assay showed that CCL5 protein enhanced the migration of T cells,supporting that CCL5 is crucial for CD8 ＋ T cells recruitment in vivo.Intriguingly,CCL5 expression was down-regulated in advanced patients （stage Ⅱb-Ⅳ）.The accumulation of CD8 ＋ T cells and CCR5 ＋ CD8 ＋ T cells was strongly reduced in advanced patients,suggesting

关 键 词：食管肿瘤 CCL5 T淋巴细胞 肿瘤浸润性淋巴细胞 单核细胞 穿孔素 颗粒酶B 

分 类 号：R735.1[医药卫生—肿瘤]