基于流式细胞技术分析固有淋巴细胞亚群失衡与非小细胞肺癌的关系

Analysis of relationship between innate lymphocyte subsets inbalance and NSCLC based on flow cytometry technology

摘要:

目的：探讨固有淋巴细胞（innate lymphoid cells，ILCs）亚群与非小细胞肺癌（non-small cell lung cancer，NSCLC）发生发展及预后的关系。方法：选取2012年1月至2014年1月安徽医科大学第一附属医院收治的70例均经病理学诊断为NSCLC的患者为NSCLC组，选择同期来本院的性别比、年龄相匹配的70例健康体检者为对照。采用流式细胞技术检测２组ILCs亚群比例，采用酶联免疫吸附法检测ILCs亚群相关细胞因子水平，收集患者临床资料与随访资料，分析ILC第一亚群（innate lym－phoid cell type 1，ILC1）、ILC第二亚群（innate lymphoid cell type 2，ILC2）、ILC第三亚群（innate lymphoid cell type 3，ILC3）与NSCLC临床特征、预后的关系。结果：与对照组相比，NSCLC组ILC1及干扰素-γ（interferon-γ，IFN-γ）水平分别为（8.61±3.78）%和（27.24±12.85） ng/L，明显降低（P＜0.05），ILC2、ILC3、白细胞介素-5（interleukin-5，IL-5）和白细胞介素-17（inter－leukin-17，IL-17）水平分别为（1.29±0.64）%、（3.64±1.93）%、（22.70±9.94） ng/L和（8.10±2.09） ng/L，明显升高（P＜0.05）。ILC1与IFN-γ呈明显正相关（r=0.780，P＜0.05），ILC2与IL-5呈明显正相关（r=0.704，P＜0.05），ILC3与IL-17呈明显正相关（r=0.654，P＜0.05）；ILC1、ILC2及ILC3水平均与淋巴结转移及TNM分期有关（P＜0.05）。无淋巴结转移、TNM分期较低、ILC1水平≥8.98%、ILC2水平＜1.37%的患者总生存期（overall survival，OS）明显较长（P＜0.05）。ILC1≥8.98%的患者中位生存时间为49个月，ILC1＜8.98%的患者中位生存时间为38个月，2组生存曲线有明显差异（P＜0.05）。ILC2≥1.37 %的患者中位生存时间为34个月，ILC2＜1.37%的患者中位生存时间为46个月，2组生存曲线有明显差异（P＜0.05）。多因素分析结果显示，淋巴结转移，TNM Ⅲ、Ⅳ分期，ILC2≥1.37%是OS的独立危险因素（P＜0.05），ILC1≥8.98%是OS的独立保护因素（P＜0.05）。结论： NSCLC患者表现为ILCs亚群失衡，ILC2、ILC3亚群具有明显优势，检测ILC1、ILC2亚群水平可在一定程度上评估患者疾病发展与预后情况。

Abstract:

Objective：To explore the relationship between the innate lymphoid cells（ILCs） and the occurrence，development and prognosis of non-small cell lung cancer（NSCLC）. Methods：70 patients diagnosed as NSCLC from January 2012 to January 2014 in the First Affiliated Hospital of Anhui Medical University were enrolled as NSCLC group. 70 healthy examiners with matching gender ratio and age in the hospital for physical examination at the same time period were selected as control group. Flow cytometry was used to detect the proportion of ILCs subsets in the two groups. The levels of ILCs-related cytokines were detected by enzyme-linked immunosorbent assay（ELISA）. The clinical data and follow-up data were collected，and the relationship between ILC1，ILC2 and ILC3 and clinical features and prognosis of NSCLC was analyzed. Results：Compared with control group，the levels of ILC1 and IFN-γ with （8.61±3.78）% and （27.24±12.85） ng/L in NSCLC group decreased（P<0.05），while the levels of ILC2，ILC3，IL-5 and IL-17 with （1.29±0.64）%，（3.64±1.93）%，（22.70±9.94） ng/L and （8.10±2.09） ng/L increased（P<0.05）. ILC1 was positively cor－related with IFN-γ（r=0.780，P<0.05），ILC2 was positively correlated with IL-5（r=0.704，P<0.05），and ILC3 was positively correlated with IL-17（r=0.654，P<0.05）. The levels of ILC1，ILC2 and ILC3 were associated with lymph node metastasis and TNM staging（P<0.05）. The overall survival（OS） was significantly longer among patients with no lymph node metastasis，low TNM stage，ILC1 level ≥8.98% and ILC2 level <1.37%（P<0.05）. The median survival time was 49 months in patients with ILC1≥8.98%，and 38 months in patients with with ILC1<8.98%，and there were significant differences in the survival curves between the two groups（P<0.05）. The median survival time was 34 months in patients with ILC2≥1.37% and 46 months in patients with ILC2<1.37%，and there were significant differences in the survival curves between the two groups（P<0.05）. Multivariate analysis showed that lymph node metastasis，TNM stage III，IV and ILC2≥1.37% were indepen－dent risk factors for OS（P<0.05），and ILC1≥8.98% was an independent protective factor for OS（P<0.05）. Conclusion：NSCLC patients have an imbalance of ILCs，and ILC2 and ILC3 have obvious advantages. The detection of ILC1 and ILC2 can evaluate the development and prognosis of disease to a certain extent.