血管内皮钙黏蛋白在非小细胞肺癌血管生成中的作用  

Effect of vascular endothelial cadherin on angiogenesis of non-small cell lung cancer

作　　者：陶韬[1] 陈虹[1] 顾雪霜[1] 周俊豪[1] 彭单伊[1] 张丽君[1] 

机构地区：[1]重庆医科大学附属第一医院呼吸内科,重庆400016

出　　处：《吉林大学学报（医学版）》2013年第3期559-564,共6页Journal of Jilin University：Medicine Edition

基　　金：重庆市自然科学基金计划项目资助课题(cstc2011jjA10052)

摘　　要：目的:探讨血管内皮钙黏蛋白(VE-cadherin)在非小细胞肺癌(NSCLC)血管生成中的作用,为NSCLC的抗血管生成治疗提供实验依据。方法:将人肺癌A549细胞分为空白对照组、干扰组和阴性对照组,VE-cadherin-siRNA瞬时转染A549细胞,实时定量PCR和Western blotting法检测VE-cadherin在各组细胞中表达水平;新型四唑氮盐(MTS)比色法检测A549细胞吸光度(A490)值的变化;流式细胞术检测细胞周期和凋亡率;ELISA法分析A549细胞瞬时转染上清中VE-cadherin的表达水平;并分析上清液作用后人脐静脉内皮细胞(HUVECs)A490值、各周期(G0/G1、G2/M和S)细胞比率、凋亡率和Matrigel小管形成数的改变。结果:干扰组VE-cadherin mRNA的表达水平(0.299±0.039)明显低于空白对照组(1.137±0.082)和阴性对照组(1.001±0.076)(P<0.05);干扰组VE-cadherin蛋白的表达水平(0.297±0.045)明显低于空白对照组(0.833±0.119)和阴性对照组(0.794±0.095)(P<0.05);各组A549细胞A490值、各周期细胞比率和凋亡率比较差异无统计学意义;干扰组上清液中VE-cadherin的表达水平[(2.89±0.07)μg.L-1]明显低于空白对照组[(11.43±0.09)μg.L-1]和阴性对照组[(11.15±0.04)μg.L-1](P<0.05)。干扰组与阴性对照组上清液作用后G0/G1、G2/M和S期HUVECs所占百分比比较差异无统计学意义(P>0.05);干扰组上清液明显抑制了HUVECs的增殖,作用24、48和72h细胞增殖抑制率分别为29.2%、33.8%和30.1%;作用48h干扰组HUVECs的凋亡率为27.12%±5.22%,显著高于阴性对照组(13.43%±3.50%)(P<0.05)。干扰组HUVECs小管形成数为1.53±0.31,显著低于阴性对照组(14.53±1.51)(P<0.05)。结论:VE-cadherin可能通过促进血管生成参与NSCLC的发生发展,有望成为NSCLC抗血管生成治疗的新靶点。Objective To investigate the role of vascular endothelial cadherin（VE-cadherin） on the angiogenesis of non-small cell lung cancer（NSCLC）,and to provide experimental basis for anti-angiogenic therapy of NSCLC.Methods The A549 cells were divided into blank control group,interference group and negative control group.VE-cadherin-siRNA was transiently transfected into A549 cells,and Real-time quantitative PCR and Western blotting methods were used to detect the expression levels of VE-cadherin mRNA;the changs of cell absorbance（A490） value was detected by four methy1 thiazolyl tetrazolium（MTS） assay;the cell cycle and apoptosis were detected by flow cytometry;the expression level of VE-cadherin in transiently transfected A549 cells supernatant was detected by ELISA method;the changes of the A490 value,ratios of cell cycle（G0/G1,G2/M and S）,apoptotic rate and Matrigel tubule formation number of human umbilical vein endothelial cells（HUVECs） were analyzed after treated with supernatant.Results The expression level of VE-cadherin mRNA in interference group（0.299±0.039） was significantly lower than those in blank control group（1.137±0.082） and negative control group（1.001±0.076）（P0.05）;the expression level of VE-cadherin protein in interference group（0.297±0.045） was significantly lower than those in blank control group（0.833±0.119） and negative control group（0.794±0.095）（P0.05）.The A490 values,cell cycle ratios and apoptotic rates of the A549 cells in blank control group,interference group and negative control group had no significant differences（P0.05）;the expression level of VE-cadherin in the supernatant in interference group was（2.89±0.07）μg·L-1,which was significantly lower than those in blank control group [（11.43±0.09） μg·L-1] and negative control group [（11.15±0.04） μg·L-1]（P0.05）.Between interference group and negative group,the percentages of HUVECs at G0/G1,G2/M and S phases had no significant difference after treated wi

关 键 词：血管内皮钙黏蛋白 血管生成 癌 非小细胞肺 人脐静脉内皮细胞 

分 类 号：R734.2[医药卫生—肿瘤]