自分泌VEGF/VEGFR环路在NK/TCL中的表达及其对肿瘤迁移和侵袭的调控  

Autocrine loop of VEGF/VEGFR signaling promotes cell migration and invasion in NK/T-cell lymphoma cells

作　　者：董一文[1] 金震[1] 丁浩[2] 常君[1] 张文皓[1] 李清[3] 曹祥山[4] 王立峰[5] 陶荣[1] 

机构地区：[1]上海交通大学医学院附属新华医院血液科,上海200092 [2]复旦大学附属眼耳鼻科医院放疗科 [3]常州市第一人民医院病理科 [4]常州市第一人民医院血液科 [5]上海交通大学医学院附属新华医院病理科

出　　处：《临床血液学杂志》2014年第3期379-385,共7页Journal of Clinical Hematology

摘　　要：目的:检测VEGF/VEGFR自分泌环路在NK/T细胞淋巴瘤(NK/TCL)细胞株中的表达,探讨VEGF/VEGFR通路对NK/TCL细胞迁移和侵袭的影响及机制。方法:利用RT-PCR及免疫荧光技术检测细胞VEGF及VEGFR的表达,蛋白印迹技术检测KDR的磷酸化。运用transwell技术观察VEGF/VEGFR自分泌环路经下游信号转导通路对细胞迁移和侵袭的影响。ELISA检测VEGF/VEGFR自分泌环对细胞生成基质金属蛋白酶(MMP)-2和MMP-9的影响,探讨MMP活性与细胞侵袭力的关系。结果:三株NK/TCL细胞株SNK-1、SNK-6和SNT-8均同时表达VEGFA及其受体FLT1和KDR,形成肿瘤细胞特异的VEGF/VEGFR自分泌环路,并可诱导KDR磷酸化。VEGF/VEGFR自分泌环路显著促进NK/TCL的细胞迁移能力,可能由受体后AKT和p38信号通路调控,并可通过促进MMP-2和MMP-9的分泌促进细胞的侵袭能力。结论:NK/TCL中存在肿瘤细胞特异的VEGF/VEGFR自分泌环路,具有促进肿瘤细胞的迁移和侵袭能力的作用,可能为临床治疗的潜在靶点。Objective：To identify the expression pattern of VEGF and VEGFRs in NK/T-cell lymphoma（NK/ TCL）cell lines and investigate the role of VEGF/VEGFR signaling in regulation of cell migration and invasion. Method：RT-PCR and immunofluorescence were performed to identify the expression of VEGF and VEGFRs. Western blot was used to detect the levels of phosphorylation of KDR.Transwell assay was applied to determine the rates of cell migration and invasion induced by VEGF/VEGFR signaling and activation of its downstream kinases.Concentrations of MMP-2and MMP-9were quantified by ELISA.Result：Co-expression of VEGFA with its receptors FLT1and KDR were identified in three NK/TCL cell lines SNK-1,SNK-6and SNT-8.Significant phosphorylation of KDR was detected in all these cell lines,which forms an autocrine loop of VEGF/VEGFR signaling. This signaling promoted the rates of cell migration and invasion.Inhibition of AKT and p38,the key downstream kinases of VEGF/VEGFR signaling,significantly reduced the rates of cell migration.VEGF/VEGFR signaling induced secretion of high levels of MMP-2and MMP-9,which involved in cell invasion.Conclusion：An autocrine VEGF/VEGFR loop is aberrantly expressed on NK/TCL cells.Autocrine VEGF signaling promotes lymphoma cells migration and invasion and may be a potential therapeutic target.Cell migration and invasion,which appears to be a potential therapeutic target in NK/TCL.

关 键 词：NK T淋巴瘤 血管内皮生长因子 自分泌 细胞迁移 细胞侵袭 

分 类 号：R733.4[医药卫生—肿瘤]