摘要
目的探讨肝细胞生长因子（HGF）及其受体c-Met对大鼠骨髓内皮祖细胞（EPCs）迁移能力的影响及其机制。方法提取、培养和鉴定大鼠骨髓EPCs;重组腺病毒载体介导c-Met基因转染EPCs,用real-time PCR、Western blot和CCK8分别检测c-Met的表达和细胞增殖;分别用0、5、10、20和50 ng/m L HGF处理Ad-c-Met-EPCs,并通过Transwell迁移系统检测Ad-c-Met-EPCs的迁移能力;选取适宜浓度的HGF处理EPCs、Ad-GFP-EPCs和Ad-c-MetEPCs,并设置PBS对照组和磷脂酰肌醇3-激酶抑制剂LY294002组（同时加入HGF和10 g/L的LY294002）,通过Transwell迁移系统和Western blot分别检测各组细胞的迁移能力、Akt和P-Akt的表达。结果 1）Ad-c-Met-EPCs中c-Met基因与蛋白均为高表达（P〈0.05）。2）c-Met基因对EPCs的增殖无明显影响。3）HGF在0～50 ng/m L范围内呈浓度依赖性促进Ad-c-Met-EPCs迁移。4）HGF＋Ad-c-Met-EPCs组的迁移能力和P-Akt的表达显著高于对照组、HGF＋EPCs组、HGF＋Ad-GFP-EPCs组和HGF＋LY294002＋Ad-c-Met-EPCs组（P〈0.05）。结论 HGF/c-Met能够显著提高EPCs的迁移能力;HGF/c-Met可能通过PI3K/Akt信号通路促进EPCs的迁移。
Objective To investigate the effect of hepatocyte growth factor（ HGF） and its receptor c-Met on the migration ability of rat bone marrow-derived endothelial progenitor cells（ EPCs） and its mechanism. Methods The EPCs were separated,obtained and identified. The Ad-c-Met-EPCs were obtained by recombinant adenovirus vector mediates c-Met transfered EPCs. The expression of c-Met and proliferation capacity of each group were detected by real-time PCR,Western blot,and CCK8 respectively. The migration ability of Ad-c-Met-EPCs with different concentrations of HGF was checked by Transwell system. The test cells were processed by proper concentration of HGF,while the control group and phosphatidylinositol 3-kinase inhibitor group（ add HGF and 10 g/L LY294002） were also setted. The migration ability,Akt and P-Akt of the each group cells were detected by Tran-swell system and Western blot. Results 1） The results of q PCR and Western blot showed that c-Met gene and protein in Ad-c-Met-EPCs had a high expression（ P〈 0. 05）. 2） The c-Met gene did not have significant impact on the appreciation capacity of EPCs. 3） HGF increased Ad-c-Met-EPCs migration with concentration increasing（ 0-50 ng/m L）. 4） The migration capacity of HGF ＋ Ad-c-Met-EPCs and P-Akt protein were significantly higher than other groups（ P〈 0. 05）. Conclusions HGF/c-Met can remarkably increase the migration capacity of EPCs. Besides,HGF/c-Met may accelerate the migration of EPCs through PI3 K/Akt signal path.
出处
《基础医学与临床》
CSCD
2016年第12期1611-1617,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(81370432)
重庆市教委科学技术研究项目(KJ1500238)
重庆市自然科学基金(cstc2011jjA10096)
国家临床重点专科建设项目(国卫办医函[2013]544)
关键词
HGF
C-MET
内皮祖细胞
细胞迁移
hepatocyte growth factor
c-Met
endothelial progenitor cells
cell migration
