自分泌PDGF/PDGFR环路对NK/T细胞淋巴瘤细胞增殖的影响及其机制研究  

Autocrine PDGF/PDGFR loop in the proliferation of NK/T-cell lymphoma cells

作　　者：金震[1] 李青[2] 丁浩[3] 常君[1] 张文皓[1] 王立峰[4] 曹祥山[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,200092 [2]江苏省常州市第一人民医院病理科 [3]复旦大学附属眼耳鼻科医院放疗科 [4]上海交通大学医学院附属新华医院病理科,200092 [5]江苏省常州市第一人民医院血液科

出　　处：《白血病．淋巴瘤》2014年第6期325-330,共6页Journal of Leukemia & Lymphoma

摘　　要：目的 了解PDGF及PDGFR在NK/T细胞淋巴瘤细胞株中的表达,观察PDGF/PDGFR通路对NK/T细胞淋巴瘤细胞增殖与凋亡的影响及机制.方法 利用免疫荧光检测来源于NK/T细胞淋巴瘤患者的SNK-1及SNK-6肿瘤细胞PDGF及PDGFR的表达,通过细胞计数观察干预PDGF/PDGFR通路活性对细胞增殖的影响,流式细胞术检测细胞周期及细胞凋亡的变化,蛋白质免疫印迹法检测PDGFR及其下游激酶ERK1/2和AKT的磷酸化和细胞周期调节蛋白的表达.结果 细胞株SNK-1及SNK-6中存在PDGFA、PDGFB、PDGFRA、PDGFRB的共表达,自分泌的PDGF能诱导PDGFR磷酸化并活化AKT及ERK1/2信号转导,促进细胞增殖,而伊马替尼抑制PDGFR磷酸化后显著抑制了细胞增殖,伊马替尼作用第6天SNK-1及SNK-6细胞的生长抑制率分别达到（42.7±3.3）％和（36.7±3.8）％,与对照组间差异具有统计学意义（均P＜ 0.01）,但未显著增加细胞凋亡,细胞周期由G0/G1期向S期的进展发生阻滞,SNK-1和SNK-6细胞S期的比例分别为（29.2±2.1）％和（37.5±2.4）％,与对照组的（37.3±2.0）％和（42.8±2.3）％相比差异具有统计学意义（均P＜ 0.05）,伴有p21和p27表达的显著增加.结论 NK/T细胞淋巴瘤细胞中存在异常表达的自分泌PDGF/PDGFR环路,该通路具有信号转导活性,能促进细胞增殖及细胞周期进展.Objective To identify the expression pattern of PDGF/PDGFR in NK/T-cell lymphoma cell lines and investigate the effect and underlying mechanism of PDGF/PDGFR signaling on cell proliferation and apoptosis in NK/T-cell lymphoma.Methods Immunofluorescence was performed to identify the expression of PDGF and PDGFR.Cell proliferation was reflected by direct cell counting.Flow cytometry was used to determine the distribution of cell cycle and cell apoptosis.Western blot was conducted to determine the phosphorylation of PDGFR and downstream kinases ERK1/2 and AKT,and expression levels of cell cycle regulating proteins.Results Co-expression of PDGFA and PDGFB with PDGFRA and PDGFRB were detected on both cell lines SNK-1 and SNK-6.Phosphorylation of PDGFR and activation of downstream kinases ERK1/2 and AKT was induced by the autocrine PDGF signaling.Inhibition of PDGFR phosphorylation by imatinib reduced proliferation of both cells without inducing significant cell apoptosis.The inhibition rate of SNK-1 and SNK-6 was （42.7±3.3） % and （36.7±3.8） %,respectively,when treated with Imatinib for 6 days,compared with control group,there was statistical significance （P 〈 0.01）.Analysis of cell cycle distribution showed that cells were accumulated at G0/G1 phase with increased expression of p21 and p27,which indicated the cells were arrested at transition from G0/G1 to S phase.The proportion of cells in S phase in SNK-1 and SNK-6 was decreased from （37.3±2.0） % and （42.8±2.3） % to （29.2±2.1） % and （37.5± 2.4） %,respectively.The difference was statistically significant （P 〈 0.05）.Conclusion Autocrine PDGF/PDGFR loop was aberrantly expressed on NK/T-cell lymphoma cells.Autocrine PDGF could induce PDGFR phosphorylation and activate downstream signaling,thereby promoting cell cycle progression and increasing cell proliferation.

关 键 词：淋巴瘤 结外NK-T细胞 白分泌 细胞增殖 细胞周期 血小板源生长因子 

分 类 号：R733.1[医药卫生—肿瘤]