文章摘要
张红梅,徐娟,娄鉴芳,史新惠,黄蕾,孙瑞红,张淑平,柯星,潘世扬,王芳.卵巢癌患者外周血单个核细胞Toll样受体2的表达及其诱导的IL-10表达[J].南京医科大学学报,2014,(4):490~494
卵巢癌患者外周血单个核细胞Toll样受体2的表达及其诱导的IL-10表达
IL-10 response in peripheral blood mononuclear cells of ovarian cancer patients through Toll-like receptor-2 pathways
投稿时间:2013-07-11  
DOI:10.7655/NYDXBNS20140418
中文关键词: 卵巢癌  Toll样受体2  外周血单个核细胞  IL-10
英文关键词: ovarian cancer  Toll like receptor 2  PBMC  IL-10
基金项目:国家自然科学基金(30901344,81272324);江苏省实验诊断学重点实验室基金(XK201114)
作者单位
张红梅 南京医科大学第一附属医院检验学部,江苏南京210029
南京医科大学附属江宁医院输血科,江苏 南京211100 
徐娟 南京医科大学第一附属医院检验学部,江苏南京210029 
娄鉴芳 南京医科大学第一附属医院检验学部,江苏南京210029 
史新惠 南京医科大学第一附属医院检验学部,江苏南京210029 
黄蕾 南京医科大学第一附属医院检验学部,江苏南京210029 
孙瑞红 南京医科大学第一附属医院检验学部,江苏南京210029 
张淑平 南京医科大学第一附属医院检验学部,江苏南京210029 
柯星 南京医科大学第一附属医院检验学部,江苏南京210029 
潘世扬 南京医科大学第一附属医院检验学部,江苏南京210029 
王芳 南京医科大学第一附属医院检验学部,江苏南京210029 
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中文摘要:
      目的:观察Toll样受体2(Toll-like receptors 2,TLR2)在卵巢癌患者外周血单个核细胞(peripheral blood mononuclear cells,PBMC)中的表达及其诱导IL-10的表达水平,初步探索TLR2在卵巢癌发生发展中的作用机制?方法:收集20例卵巢癌患者?20例妇科良性疾病患者及20例同期健康体检女性EDTA抗凝外周全血,采用荧光定量PCR技术检测3组PBMC中TLR2的表达水平,并比较3组表达差异情况?TLR1?TLR2和TLR6相应配体刺激PBMC,细胞中细胞因子IL-10表达水平采用荧光定量PCR技术进行检测;流式细胞技术(FACS)检测各组IL-10分泌水平?结果:各组PBMC中TLR2均有表达;卵巢癌组PBMC 中TLR2表达量显著高于良性疾病组和健康对照组(P < 0.05),良性疾病组TLR2表达水平与健康对照组间无显著性差异;各组PBMC经TLR2的配体HKLM刺激24 h后,卵巢癌组IL-10 mRNA表达明显增强,分别为良性疾病组和健康对照组的2.25?2.33倍,妇科良性疾病组IL-10 mRNA表达水平与健康对照组间无显著差异;TLR6配体FSL-1刺激后,卵巢癌组IL-10 mRNA表达水平分别为良性疾病组和健康对照组的1.95?2.16倍,差异具有统计学意义(P < 0.05),而良性疾病组与健康对照组间无显著差异?FACS结果显示各组PBMC经TLR1的配体Pam3CSK4刺激24 h后,卵巢癌组?良性疾病组及健康对照组IL-10分泌水平中位值(M)分别为46.70?61.53?31.11 pg/ml,差异无统计学意义;经TLR2配体HKLM刺激24 h后,卵巢癌组IL-10分泌水平(M = 150.46 pg/ml)显著高于良性疾病组(M = 38.86 pg/ml)及健康对照组(M = 44.93 pg/ml),差异有统计学意义(P < 0.05),良性疾病组与健康对照组间无显著性改变;TLR6配体FSL-1刺激24 h后,卵巢癌组?良性疾病组及健康对照组IL-10分泌水平中位值分别为20.20?31.12?35.48 pg/ml,3组间无显著差异?结论:卵巢癌患者PBMC中TLR2表达水平显著增高,经其介导的细胞因子IL-10表达水平的改变可能与卵巢癌的免疫抑制相关,在卵巢癌的肿瘤免疫逃逸中发挥重要作用?
英文摘要:
      Objective:To investigate the expression of Toll-like receptors 2(TLR2) in peripheral blood mononuclear cells(PBMCs)from patients with ovarian cancer,and its role in inducing the expression of IL-10. Methods:We collected PBMCs from 20 patients with ovarian cancer, 20 with benign diseases and 20 healthy females. Expression levels of TLR2 mRNA in PBMCs in the 3 groups were determined by real-time quantitative PCR and then compared. PBMCs were then stimulated with TLR1,TLR2 and TLR6 ligands. The expression and secretion levels of IL-10 in each troup were assessed by real-time PCR and FACS,respectively. Results:TLR2 were all expressed in PBMCs of the three groups, and the expression levels of TLR2 mRNA in PBMCs in patients with ovarian cancer were higher than the benign group and the healthy controls(both P < 0.05). There was no significant difference in TLR2 between the benign group and the healthy controls. After stimulated with HKLM(TLR2 ligand) for 24 h,IL-10 mRNA level was significantly higher in the ovarian cancer group compared to those in the benign controls and the healthy controls(Fold = 2.25,P < 0.05;Fold = 2.33,P < 0.05). There was no significant difference in IL-10 mRNA level between the benign controls and the healthy controls. Increased expression of IL-10 was also observed upon stimulation by FSL-1(TLR6 ligand) in ovarian cancer patients compared to those in the benign controls and the healthy controls(Fold = 1.95,P < 0.05;Fold = 2.16,P < 0.05). No signifiant difference was found between the benign controls and the healthy controls. Furthermore,there was an observable increased(P < 0.05) IL-10 secretion level upon stimulation by TLR2 ligand HKLM for 24 h in ovarian cancer patients(Medium = 150.46) compared to the benign controls(Medium= 38.86 pg/ml)and the healthy controls(Medium = 44.93 pg/ml),while no significant difference was identified between the benign group and the healthy controls. It was particularly noteworthy that PBMCs did not show significant up-regulation of IL-10 production in response to Pam3CSK4(TLR1 ligand) (M=46.70,61.53 and 31.11 pg/ml,respectively) and FSL-1 stimulation (M=20.20,31.12 and 35.45 pg/ml,respectively) in the 3 groups. Conclusion:TLR2 was highly expressed in PBMCs in ovarian cancer patients. IL-10 may be related to the immune evasion of ovarian cancer,which promotes the tumor progression by tumor immune escape.
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