Detection and Quantification of Th17 Cells and IL-17 in the Peripheral Blood and Tumor Tissue in Patients with Non-small Cell Lung Cancer
|Keywords||Th17 cell IL-17 NSCLC|
【Objective】To investigate Th17 cell quantification and the concentrations of cytokine IL-17 in patients with non-small cell lung cancer(NSCLC), and to explore the possible pathological mechanism of these cells involved in the development of NSCLC.【Methods】Eighty-three patients with newly diagnosed non-small cell lung cancer (NSCLC) were involved in the study from July 2010 to January 2011, and 20 healthy volunteers were recruited as normal controls (NCs). Th1 and Th17 cells in blood samples from NSCLC patients were evaluated with flow cytometry (FCM), and the concentrations of cytokine IL-17 in serum were measured by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry(IHC) was used to determine IL-17 expression in paraffin-embedded tissues with NSCLC, and the mRNA expression levels of IL-17 in tumor tissues were investigated by real-time fluorescence quantitative PCR (real-time PCR). The potential interrelationship of Th17 cells, IL-17~+cells or IL-17 and the clinical parameters of NSCLC patients were analyzed.【Results】1. Compared with normal controls, patients with NSCLC showed lower quantification of Th1 cells (5.09±2.52% vs 6.83±2.02%, P<0.05) and higher quantification of Th17 cells and IL-17 concentrations(1.59±0.97% vs 0.69±0.41%, P<0.05), (8.13±4.89 pg/ml vs 4.92±2.20 pg/ml, P<0.05).2. The quantification of Th17 cells and IL-17 concentrations had postive correlation with the clinical stage, patients with advanced disease(stageⅢ-Ⅳ) were higher than early disease(stageⅠ-Ⅱ) (1.73±1.04% vs 1.22±0.79%, P<0.05), (8.13±4.89pg/ml vs 5.30±3.33pg/ml, P=0.054). For further analysis, IL-17 concentrations were significantly higher in patients with advanced disease (stageⅢ-Ⅳ) than those in normal controls(8.13±4.89 pg/ml vs 4.92±2.20 pg/ml, P<0.05).3. Th17 cell quantification in NSCLC patients show positive correlation with IL-17 concentrations(r=0.20, P=0.22), but the difference was not statistically significant.4. Compared with peritumor tissues, intratumor tissues showed higher expression of IL-17~+cells and IL-17mRNA((16/35)% vs (0/10)%, P<0.01), (0.0284±0.0066 vs 0.0029±0.0009, P <0.01).5. The expression of IL-17~+ cells in intratumor tissues had postive correlation with the clinical stage and mediastinal lymph node, patients with advanced disease (stageⅢ) were higher than those in early disease(stageⅠ-Ⅱ)((9/13)% vs (7/22)%, P<0.05), and patients with positive mediastinal lymph node(N2) were higher than those with negative(N0-1) ((8/10)% vs (8/25)%, P<0.05). The expression of IL-17mRNA in patients with advanced disease (stageⅢ)were higher than those in early disease(stageⅠ-Ⅱ) (0.0399±0.0121 vs 0.0191±0.0058, P=0.087)6. The IL-17~+ cells per HPF in intratumor tissues show positive correlation with the levels of IL-17mRNA (r=0.64, P<0.01).【Conclusion】There are higher quantification of Th17 cells and higher expression of IL-17 in the peripheral blood and intratumor tissue in patients with NSCLC and they have postive correlation with the clinical stage. Detecting the quantification of Th17 cells and expression of IL-17 would contribute to prediction of the disease progression and prognosis.