酪氨酸激酶受体B-脑源性神经营养因子信号传导通路对神经母细胞瘤细胞分泌血管内皮生长因子及基质金属蛋白酶-9的影响  

Effects of tyrosine kinase receptor B-brain-derived neurotrophic factor signal pathway on the secretion of vascular endothelial growth factor and matrix metalloproteinase-9 of neuroblastoma

作　　者：刘建英[1] 高惠敏 李爱敏[1] 蔡维艳[1] 初清[1] 

机构地区：[1]青岛大学医学院附属烟台毓璜顶医院儿科,264000 [2]烟台市中医院,264000

出　　处：《中国小儿急救医学》2013年第4期398-402,共5页Chinese Pediatric Emergency Medicine

摘　　要：目的探讨酪氨酸激酶受体B（tyrosine kinase receptorB，TrkB）-脑源性神经营养因子（brain—derived neurotrophic factor，BDNF）信号传导通路对神经母细胞瘤（neuroblastoma，NB）细胞SH-SY5Y分泌血管内皮生长因子（vascular endothelial growth factor，VEGF）及基质金属蛋白酶-9（matrix metalloproteinase-9，MMP-9）的影响。方法用全反式维甲酸（all—trans retinoic acid，ATRA）诱导SH—SY5Y细胞表达TrkB，加入外源性BDNF，从而激活TrkB—BDNF信号传导通路及其3条下游信号通路。再用特异性酪氨酸激酶抑制剂K252a阻断TrkB—BDNF信号传导通路；用磷脂酰肌醇-3激酶（phos—phatidylinositol 3-hydroxy kinase，P13K）抑制剂LY294002、磷脂酶C抑制剂U73122、丝裂原活化蛋白激酶抑制剂U0126分别阻断TrkB-BDNF的3条相应下游信号传导通路。采用酶联免疫吸附法检测细胞培养上清液中VEGF及MMP-9含量。结果ATRA＋BDNF组VEGF[（485．89±109．99）pg／ml]及MMP-9[（15．73±1．72）pg／ml]含量明显高于对照组及ATRA组（P〈0．05）；ATRA＋BDNF＋K252a组VEGF[（272．42±86．33）pg／ml]及MMP-9[（5．25±1．44）pg／ml]含量明显低于ATRA＋BDNF组（P〈0．05），与对照组及ATRA组比较差异无统计学意义（P〉0．05）；ATRA＋BDNF＋LY294002组VEGF[（314．12±24．68）pg／ml]及MMP-9[（4．91±1．08）pg／ml]含量明显低于ATRA＋BDNF组（P〈0．05），与对照组及ATRA组比较差异无统计学意义（P〉0．05）；ATRA＋BDNF＋U73122组VEGF[（444．08±64．49）pg／ml]及MMP-9[（13．28±3．38）pg／ml]含量与ATRA＋BDNF组比较差异无统计学意义（P〉0．05）。ATRA＋BDNF＋U0126组VEGF[（429．97±19．95）pg／ml]及MMP-9{（13．96±4．45）pg／ml]含量与ATRA＋BDNF组比较差异无统计学意义（P〉0．05）。结论激活TrkB-BDNF信号传导通路可促进NB细胞合成、分泌VEGF及MMP-9。TrkB—BDNF信号传导通路可能通过进一步激活其下游P13K／Akt�Objective To study the effects of tyrosine kinase receptor B-brain-derived neurotrophic factor （TrkB-BDNF） signal pathway on the secretion of vascular endothelial growth factor （VEGF） and ma- trix metalloproteinases-9 （MMP-9） of neuroblastoma. Methods We used ail-trans retinoic acid （ATRA） to induce the high expression of TrkB in the SH-SY5Y cell line, and then added the ectogenid BDNF to activate the TrkB-BDNF and its three downstream signal pathways. TrkB-BDNF signal pathway was inhibited by spe-cific tyrosine kinase inhibitor K252a. The three downstream signal pathway was respectively inhibited by LY294002 （the phosphatidylinositol 3-hydroxy kinase （PI3K） pathway inhibitor） ,U73122（ the phospholipase C pathway inhibitor） ,U0126 （the mitogen activated protein kinase pathway inhibitor）. Enzyme linked immu-nosorbent assay was used to detect the concentration of VEGF and MMP-9 protein in the SY5Y cell culture supematants. Results VEGF [ （ 485. 89 ±109. 99 ） pg/ml] and MMP-9 E（15.73 ±1.72） pg/ml] protein levels in neuroblastoma cells cultured in serum-free media in the group of ATRA ＋ BDNF were significantly higher than that of the control group and ATRA alone group（ P 〈 0. 05 ）. VEGF [ （272.42 ± 86. 33 ） pg/ml ] and MMP-9 [ （5.25 ± 1.44） pg/ml] protein levels in the group of ATRA ＋ BDNF ＋ K252a were significant- ly lower than those of the ATRA ＋ BDNF group（ P 〈 0.05 ） and had no significant difference compared with the control group and the ATRA alone group（P 〉0. 05）. VEGF [ （314. 12 ± 24. 68） pg/ml] and MMP-9 [ （4. 91 ± 1.08 ） pg/ml] protein levels in the group of ATRA ＋ BDNF ＋ LY294002 were significantly lower than those of the ATRA ＋ BDNF group （ P 〈 0. 05 ） and had no significant difference compared with the con-trol group and the ATRA alone group（P 〉0. 05）. VEGF [ （444. 08 ±64. 49） pg/ml] and MMP-9 [ （ 13.28 ± 3.38） pg/ml] protein levels in neuroblastoma cells cultured in serum-free media in the

关 键 词：神经母细胞瘤 TrkB-BDNF信号传导通路 K252A LY294002 血管内皮生长因子 基质 金属蛋白酶-9 

分 类 号：R73[医药卫生—肿瘤]