摘要
目的在体外探讨血管内皮生长因子C(vascular endothelial growth factor C,VEGF-C)、趋化因子受体(Cys-X-Cys receptor4,CXCR4)及其配体基质细胞衍生因子-1(stromal cell-derived factor1,SDF-1)在喉癌Hep-2细胞株中的表达及其在喉癌细胞归巢中的作用。方法①分别用RT-PCR及Western blot检测喉癌Hep-2细胞株中CXCR4 mRNA和蛋白的表达和重组人SDF-1与CXCR4特异性拮抗剂(AMD3100)作用前后喉癌Hep-2细胞株中VEGF-CmRNA和蛋白表达水平的变化;②用甲基噻唑基四唑(MTT比色法)测定不同浓度重组人SDF-1以及AMD3100作用前后Hep-2细胞增殖率的变化;③Transwell小室迁移及体外侵袭实验检测不同浓度重组人SDF-1以及AMD3100对Hep-2细胞趋化活性的影响。结果①在mRNA以及蛋白水平,CXCR4及VEGF-C在Hep-2细胞株中均有高表达,在不同浓度重组人SDF-1(1、10、100ng/ml)作用Hep-2细胞株24h后,VEGF-C的表达明显上调,且有明显的剂量依赖性,并能被1μg/ml的CXCR4的特异性拮抗剂AMD3100阻断;②与对照组(未处理的Hep-2细胞)和10ng/mlSDF-1组比较,100ng/mlSDF-1组有明显促进喉癌细胞增殖的作用(P<0.05),并能被1μg/ml的AMD3100阻断;③与对照组和10ng/mlSDF-1组比较,100ng/mlSDF-1组同样有明显的促喉癌细胞迁移和侵袭的作用(P<0.05),同样也能被1μg/ml的AMD3100所阻断。结论 SDF-1/CXCR4生物轴与VEGF-C协同促进喉癌细胞归巢,可作为干预喉癌淋巴转移的新手段。
Objective To investigate the in vitro expressions of chamotatic factor CXCL12 [also known as stromal cell-derived factor 1 （SDF-1）],the receptor of the factor,Cys-X-Cys receptor 4 （CXCR4） and vascular endothelial growth factor C （VEGF-C） in human laryngeal carcinoma cell line Hep-2 and their roles in the homing of laryngeal carcinoma. Methods The expressions of CXCR4 mRNA and protein in Hep-2 cells was detected by RT-PCR and Western blotting,the expression of VEGF-C also was evaluated before and after incubated with SDF-1 （1,10,100 ng/ml） or the antagonist of CXCR4 AMD3100. Methythiazolyltetrazolium （MTT） assay was used to analyze the effect of different concentrations of SDF-1 on the proliferation of Hep-2 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of SDF-1 and AMD3100 on the migration and invasion of Hep-2 cells. Results 1）The CXCR4 and VEGF-C were both overexpressed at mRNA and protein level in Hep-2 cells,and the expression of VEGF-C in Hep-2 cells was up-regulated with a concentration-dependent model,which was inhibited by CXCR4 antagonist （P0.05）. 2）SDF-1（100 ng/ml） significantly enhanced the proliferation of Hep-2 cells compared with the control and 10 ng/ml SDF-1 groups （24 h：0.585±0.051 vs 0.425±0.045 and 0.428±0.039,P0.05; 48 h：0.631±0.062 vs 0.497±0.067 and 0.532±0.039,P0.01）. This enhancing effect of SDF-1 was significantly inhibited by 1 μg/ml AMD3100. 3）The levels of migration and invasion of the Hep-2 cells treated with 100 ng/ml SDF-1 were significantly higher than those in the control and 10 ng/ml SDF-1 （migration：286.7±24.7 vs 48.0±7.2 and 147.7±37.0; invasion：40.5±4.5 vs 4.0±1.0 and 17.7±2.0,P0.05）,and was strongly inhibited by 1 μg/ml AMD3100. Conclusion SDF-1/CXCR4 can in coordination with VEGF-C promote homing of laryngeal carcinoma cell line Hep-2,and may be a new pathway to prevent laryngeal carcinoma lymph node metastasis.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第12期1316-1320,共5页
Journal of Third Military Medical University
基金
重庆市卫生局科研基金(2007-2-070)
重庆市卫生局重点项目(2008-1-16)~~
