【作者】 张萍；

【导师】 刘奇；

【作者基本信息】 山东大学 ， 肿瘤学（专业学位）， 2015， 硕士

【摘要】 背景：肿瘤转移是导致癌症相关死亡的首要原因。现有研究发现存在一种快速的机制,能够在肿瘤发展的早期阶段驱动肿瘤细胞发生播散。肿瘤相关的炎症和肿瘤的发展密切相关,尤其是肿瘤浸润的中性粒细胞、淋巴细胞和巨噬细胞。本文的主要目是研究非小细胞肺癌原发灶与转移灶中炎细胞浸润水平的差异,并探寻有促进肿瘤细胞转移作用的关键炎细胞亚型。此外,我们对关键的炎细胞亚型也进行了相关临床研究。方法：纳入本项回顾性研究的患者共82人,包括初次诊断时伴有远处转移并且原发灶与转移灶一并切除的非小细胞肺癌患者41人,和另外41位与之配对的未发生转移只切除原发灶的患者。应用免疫组织化学染色检测123例肿瘤组织(包括转移组原发灶和转移灶各41例、未转移组原发灶41例)中CD66b+中性粒细胞、CD8+T淋巴细胞、CD163+巨噬细胞、E-钙黏蛋白和CXCL5的浸润和表达水平。应用Pearson卡方检验或Fisher精确检验进行肿瘤浸润中性粒细胞和外周血中性粒细胞与临床病理特征的相关性研究。应用Wilcoxon符号秩和检验比较原发灶与转移灶间浸润炎细胞的分布差异,以及转移组与未转移组原发灶间的分布差异。Spearman秩检验用于多项相关性研究。所有统计学分析中,若p<0.05则认为有统计学意义。结果：CD66b+中性粒细胞的肿瘤浸润水平在转移组患者的原发灶明显高于其转移灶(p<0.001)和未转移组患者的原发灶(p<0.001)。但是,CD8+T淋巴细胞(p=0.288,p=0.325)和CD163+巨噬细胞(p=0.726,p=0.975)并没有明显的浸润差异。此外,CD66b+中性粒细胞的肿瘤浸润水平和CXCL5的表达水平呈正相关(Spearman相关系数=0.505,p<0.001),而与E-钙黏蛋白的表达呈负相关(Spearman相关系数=-0.479,p<0.001)。同时,肿瘤浸润中性粒细胞的浸润水平和外周血中性粒细胞水平呈正相关(Spearman相关系数=-0.493,p<0.001)。进一步的研究发现,肿瘤浸润中性粒细胞(Spearman相关系数=0.415,p<0.001)和外周血中性粒细胞(Spearman相关系数=0.334,p=0.002)与阳性淋巴结转移个数均存在明显的相关性。结论：我们的研究表明CD66b+的中性粒细胞,作为关键的炎细胞亚型,主要在肿瘤的原发灶大量聚集,而以淋巴细胞和巨噬细胞为主的其他亚型炎细胞在原发灶和转移灶的浸润并没有明显差异。肿瘤浸润中性粒细胞,在一定程度上通过诱导上皮间质转化,对非小细胞肺癌患者的转移发挥了重要作用。此外,与肿瘤浸润中性粒细胞有明显相关性的外周血中性粒细胞,或许可以作为一项简单易行的非小细胞肺癌的转移监测指标。更多还原

【Abstract】 Background:Metastasis is the main cause of cancer mortality and a new insight that different, faster mechanism drove cancer cells dissemination in the early stage of tumor progression was uncovered. Cancer-related inflammation, especially tumor infiltrating neutrophils, lymphocytes and macrophages, are related to tumor progression. The main aim of this study was to characterize inflammatory cells in primary and metastases tumors of non-small-cell lung cancer (NSCLC) and explore which kind of inflammatory cells play a critical role in metastases. Additionally, the corresponding peripheral blood inflammatory cells were explored.Methods:A total of 82 NSCLC patients were studied in the retrospective study, including 41 patients with metastasis and 41 paired patients without metastasis.123 NSCLC specimens were obtained from 41 patients with metastasis (both primary tumors and metastases) and 41 patients without metastasis (only primary tumors). Immunohistochemistry was applied to detect the infiltration of CD66b+neutrophils, CD8+ T lymphocytes, CD 163+ macrophages and the expression of E-cadherin and CXCL5 in the 123 specimens. Pearson chi-square test or Fisher exact test was used to detect the relationship between the tumor-infiltrating neutrophils or peripheral neutrophils and clinicopathological features. Wilcoxon signed rank test was used for the comparison of the infiltration of inflammatory cells between the primary tumors and metastases of patients with metastasis, also for the comparison between the primary tumors of patients with metastasis and the primary tumors of patients without metastasis. Spearman rank test was applied for the study of correlation. p<0.05 was considered statistically significant.Results:The infiltration of CD66b+neutrophils in primary tumors of patients with metastasis was higher than corresponding metastases (p<0.001) and primary tumors of patients without metastasis (p<0.001). However, no significant difference was shown for CD8+ T lymphocytes (p=0.288,p=0.325) and CD 163+ macrophages (p=0.726, p=0.975). We also found that CD66b+ neutrophils infiltration was positively correlated with CXCL5 expression (Spearman’s correlation coefficient =0.505, p<.001) and negatively related to E-cadherin expression (Spearman’s correlation coefficient=-0.479, p<0.001). In addition, TIN was positively correlated with circulating neutrophils counts (Spearman’s correlation coefficient =0.493, p<0.001). Further studies showed that both TIN (Spearman’s correlation coefficient =0.415, p<0.001) and circulating neutrophils counts (Spearman’s correlation coefficient=0.334,p=0.002) were correlated with the number of positively-metastasis lymph nodes.Conclusions:Our study demonstrated that CD66b+ neutrophil, as a particular subset of inflammatory cells, mainly accumulated in primary tumors, while other subsets of immune cells including lymphocytes and macrophages were equally represented in primary tumors and metastases tumors. Neutrophils play a critical role in NSCLC metastases, partly via the ability to induce epithelial-mesenchymal transition. Furthermore, circulating neutrophils which were related to TIN could be a simple and easy monitoring indicator for metastasis of NSCLC.更多还原