摘要目的 应用体内外实验研究非小细胞肺癌( NSCLC)中白细胞介素7及其受体(IL-7/IL-7R)对调控细胞增殖和促进淋巴管形成的作用机制.方法 应用免疫组织化学SP法观察95例NSCLC组织中IL-7、IL-7R、cyclin D1和血管内皮生长因子(VEGF)-D的表达,分析IL-7/IL-7R与各因素的关系.然后应用四甲基偶氮唑盐法、流式细胞术、逆转录-聚合酶链反应( RT-PCR)、Western blot法、染色质免疫沉淀法、免疫共沉淀法及裸鼠体内移植实验研究肺癌中IL-7/IL-7R调控细胞增殖和促进淋巴管形成的作用机制.结果 IL-7( 63.2％,60/95)、IL-7R( 61.1％,58/95)、cyclin D1 (52.6％,50/95)和VEGF-D( 58.9％,56/95)在NSCLC中高表达.IL-7/IL-7R的表达与cyclin D1(P＜0.01,P＜0.01)、VEGF-D(P ＜0.01,P＜0.01)、淋巴管密度(P=0.005,p=0.013)和肿瘤临床分期(P=0.008,P=0.005)、转移(P＜0.01,P＜0.01)相关.IL-7/IL-7R能促进A549细胞生长；同时能增加cyclin D1和VEGF-D的表达；同时能促进c-Fos/c-Jun的表达和磷酸化及c-Fos/c-Jun形成异二聚体并促使c-Fos/c-Jun与cyclin D1和VEGF-D的启动子结合调控其转录；还能促进肺癌移植瘤生长及移植瘤淋巴管形成.结论 在NSCLC中,IL-7/IL-7R通过调控c-Fos/c-Jun的表达及活性进一步调控cyclin D1促进肿瘤细胞生长,调控VEGF-D促进淋巴管形成.

abstractsObjective To study the mechanism of interleukin 7/interleukin 7 receptor (IL-7/IL-7R) in promoting cell proliferation and inducing lymphangiogenesis of non-small cell lung cancer (NSCLC) in vivo and in vitro.Methods Immunohistochemical study for IL-7,IL-7R,cyclin D1 and vascular endothelial growth factor-D (VEGF-D) was carried out in NSCLC tissues from 95 patients.The relationship between IL-7/IL-7R expression and various parameters was analyzed. The mechanism of IL-7/IL-7R in promoting cell proliferation and inducing lymphangiogenesis was studied by methylthiazolyldipheuyl-tetrazolium bromide,fluorescence-activated cell sorting,reverse transcriptase-PCR,Western blot,co-immunoprecipitation, chromatin immunoprecipitation and nude mice experiments with xenograft tumors.Results IL-7 ( 63.2％,60/95 ),IL-7R ( 61.1％,58/95 ),cyclin D1 ( 52.6％,50/95) and VEGF-D (58.9％,56/95 ) showed that high level of expression in NSCLC. IL-7/IL-7R over-expression correlated with cyclin D1 expression ( P ＜ 0.01, P ＜ 0.01 ), VEGF-D expression ( P ＜0.01,P ＜0.01 ),increased lymphovascular density ( P =0.005,P =0.013 ),advanced clinical stage ( P =0.008,P =0.005 ) and presence of lymph node metastasis ( P ＜0.01,P ＜0.01 ).IL-7/IL-7R could promote proliferation of A549 cell,increase cyclin DI and VEGF-D expression,and enhance c-Fos/c-Jun expression and phosphorylation,resulting in formation of heterodimer. Furthermore, IL-7/IL-7R could induce binding of c-Fos/c-Jun to cyclin D1/VEGF-D promoters and regulate their transcription.IL-7/IL-7R could also promote proliferation and lymphangiogenesis of lung cancer xenograft tumors. Conclusions IL-7/IL-7R promotes c-Fos/c-Jun expression and activity in NSCLC. This further facilitates cyclin D1 expression and accelerates proliferation of cells and VEGF-D-induced lymphovascular formation.