吉林大学学报(医学版) ›› 2013, Vol. 39 ›› Issue (3): 559-564.doi: 10.7694/jldxyxb20130328

• 基础研究 • 上一篇    下一篇

血管内皮钙黏蛋白在非小细胞肺癌血管生成中的作用

陶韬,陈虹,顾雪霜,周俊豪,彭单伊,张丽君   

  1. 重庆医科大学附属第一医院呼吸内科,重庆 400016
  • 收稿日期:2012-10-17 出版日期:2013-05-28 发布日期:2013-07-01
  • 通讯作者: 陈 虹(Tel:023-89012688,E-mail:hopehong2003@yahoo.com.cn) E-mail:hopehong2003@yahoo.com.cn
  • 作者简介:陶 韬(1987-),女,重庆市人,在读医学硕±,主要从事肺癌的生物学行为及治疗研究。
  • 基金资助:

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

TAO Tao,CHEN Hong,GU Xue-shuang,ZHOU Jun-hao,PENG Dan-yi,ZHANG Li-jun   

  1. Department of Respiratory Medicine,First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,China
  • Received:2012-10-17 Online:2013-05-28 Published:2013-07-01

摘要: [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]明显低于空白对照组[(11.43±0.09)μg/L]和阴性对照组[(11.15±0.04)μg/L](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72 h细胞增殖抑制率分别为29.2%、33.8%和30.1%;作用48 h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。

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

Abstract: Abstract: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 A490 value,ratios of cell cycle(G0/G1,G2/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) (P<0.05);the expression level of VE-cadherin  protein in interference group(0.297±0.045)〖JP2〗 was significantly lower than those in blank  control group (0.833±0.119) and negative control group (0.794±〖JP〗0.095)(P<0.05).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(P>0.05);the expression level of VE-cadherin in the supernatant in interference group was (2.89±0.07)μg/L,which was significantly lower  than those in blank control group [(11.43±0.09) μg/L] and
 negative control group [(11.15±0.04) μg/L] (P<0.05).Between  interference group  and  negative group,the percentages of  HUVECs  at G0/G1,G2/M and S phases had no significant difference after treated with supernatant(P>0.05);but the proliferation of HUVECs  in interference group was inhibited by supernatant  significantly,the inhibitory rates of cell proliferation  24,48 and 72 h after treatment were 29.2%,33.8%,and 30.1%,respectively.48 h after treatment,the apoptotic rate of the HUVECs in  interference group  was 27.12%±5.22%,which was significantly higher than that in negative control group (13.43%±3.50%) (P<0.05).The number of tubule formation was 1.53±0.31,which was significantly lower than that in  negative control group(14.53±1.51)(P<0.05). Conclusion VE-cadherin may participate in the occurrence and development of NSCLC by promoting angiogenesis,which is expected to become a new target for  anti-angiogenic therapy of NSCLC.

Key words: vascular endothelial cadherin, angiogenesis, cancer,non-small cell lung, human umbilical vein endothelial cells

中图分类号: 

  • R734.2