【作者】 王伟；

【导师】 肖伟；

【作者基本信息】 山东大学 ， 临床医学， 2011， 硕士

【摘要】 研究背景肺癌和慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)是严重危害人类健康的两种常见病和多发病,肺癌的发病率和死亡率居恶性肿瘤首位,COPD发病则呈逐年上升趋势,已是全球第4位的致死性疾病。目前肺癌和COPD的具体发病机制未明,但研究表明两者的发病可能存在一定的联系。COPD是气道慢性炎症性疾病,肺癌也是一种和慢性炎症密切相关的疾病,因此,慢性炎症可能是这种联系的一种表现形式。研究发现,73%的男性肺癌患者和53%的女性肺癌患者同时患有COPD,且有前瞻性研究显示COPD患者罹患肺癌的风险高于非COPD者。但肺癌和COPD是性质不同的两种疾病,其慢性炎症并非完全相同,参与慢性炎症的细胞及炎症因子在数量和活性上也有相应的不同。T淋巴细胞(包括CD4+和CD8+T淋巴细胞)是参与慢性炎症的重要细胞,也是细胞免疫的主要效应细胞,可进一步分化为不同的亚群,如CD4+T淋巴细胞可分化为Th1、Th2、Th17等亚群,CD8+T淋巴细胞可分化为Tc1、Tc2等亚群,各亚群在肺癌和COPD的发病中均有重要作用,并表现出不同的数量和活性。Tc17细胞是2006年由He D等首先报道的CD8+T淋巴细胞的一个新亚群,在功能上与之前发现的Th17细胞类似,也以分泌IL-17A为特征,并可通过IL-17A等细胞因子参与慢性炎症反应和肺癌的发生发展,但COPD中Tc17细胞的变化情况尚未有相关报道。目的本研究旨在比较肺腺癌和COPD患者外周血Tc17细胞比例变化的差异。方法2009年10月至2010年12月于山东大学齐鲁医院呼吸科门诊就诊的患者中选取28例肺腺癌和21例COPD患者,选取本院健康医护人员和健康查体者42例作为健康对照组,收集他们的外周血标本。采用流式细胞术检测所有外周血标本Tc17细胞比例,采用实时定量PCR(real time-PCR)技术检测外周血单个核细胞(peripheral blood mononuclear cells,PBMC)RORct mRNA表达的水平,酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测外周血中IL-17A.TGF-β.IL-10.IL-6等细胞因子的水平。分析外周血Tcl7细胞比例和RORct mRNA的相关性。结果1.流式细胞术检测显示：肺腺癌组和COPD组患者外周血Tc17细胞比例分别为0.76±0.83%和0.38±0.37%，二者均显著高于健康对照组0.23±0.17%(分别为P<0.001；P=0.029),且肺腺癌组患者外周血Tc17细胞比例显著高于COPD组(P=0.034)；2.实时定量PCR技术结果显示：Tc17特征性转录调控因子RORct mRNA在肺腺癌组和COPD组与健康对照组相比表达均上调(p均<0.001)。3.ELISA检测结果显示：肺腺癌患者外周血IL-17A水平显著高于健康对照(P=0.039),而COPD患者外周血IL-17A水平与健康对照相比无统计学差异(P>0.05)；肺腺癌组和COPD组患者外周血TGF-β水平均显著高于健康对照组(分别为P=0.007；P=0.001),且COPD组显著高于肺肺腺癌组(P=0.032)；COPD患者外周血IL-10水平显著高于肺腺癌患者(P=0.042)，两病例组均高于健康对照组(分别为P=0.003；P<0.001)。4.相关性分析显示：肺腺癌组和COPD组患者外周血Tc 17细胞比例与PBMC中RORct mRNA水平均呈显著正相关(P=0.036，r=0.544；P=0.043,r=0.648)。结论肺腺癌和COPD患者外周血Tc17细胞比例均升高,且在肺腺癌中更明显。更多还原

【Abstract】 ObjectiveTo compare the frequency of Te17 cells in peripheral blood from patients with advanced lung adenocarcinoma and chronic obstructive pulmonary disease (COPD).MethodsThe frequency of Te17 cells was respectively detected from 28 patients with advanced lung adenocarcinoma,21 patients with COPD and 42 healthy volunteers by flow cytometry; Real time-PCR was used to dectect the level of RORct mRNA in peripheral blood mononuclear cells (PBMC); Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum concentration of TGF-β, IL-17A, IL-6 and IL-10. Analysis the correlationship between the frequency of Tc17 cells and the level of RORct mRNA.Results1.The frequency of Te17 cells from patients with andvanced lung cancer (0.76±0.83%) was significantly higher than that in patients with COPD (0.38±0.37%)(P=0.034), and both of them were significantly higher than that in healthy volunteers(0.23±0.17%)(P<0.001; P=0.029).2. The expression of RORct mRNA was also upregulated in patients with andvanced lung cancer and patients with COPD(P<0.001; P<0.001). 3. The level of IL-17A in patients with andvanced lung cancer was significantly higher than that in healthy volunteers (P=0.039),but there was no significant difference between patients with COPD and healthy volunteers; The serum concentration of TGF-βwas both significantly higher than that in healthy volunteers (P=0.007; P=0.001),and the level of IL-10 in patients with andvanced lung cancer was significantly lower than that in COPD patients.4. According to correlation analysis,the frequency of Te17 cells in patients with andvanced lung cancer and COPD patients were positively correlated with the level of RORct mRNA (P=0.036, r=0.544; P=0.043, r=0.648)ConclusionIn the peripheral blood, the frequency of Te17 cells from patients with andvanced lung cancer and COPD were significantly higher, and the frequency of Te17 cells from patients with andvanced lung cancer was higher than that in patients with COPD.更多还原