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纸质出版:2010
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张园园,胡国华,袁琨,但洋.SDF-1/CXCR4对喉癌细胞归巢的作用[J].陆军军医大学学报(原第三军医大学学报),2010,32(12):1316-1320.
Zhang Yuanyuan,Hu Guohua,Yuan Kun,Dan Yang.Effect of SDF-1/CXCR4 axis to homing of laryngeal carcinoma cell Hep-2[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(12):1316-1320.
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 receptor 4,CXCR4)及其配体基质细胞衍生因子-1(stromal cell-derived factor 1,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-C mRNA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、100 ng/ml)作用Hep-2细胞株24 h后,VEGF-C的表达明显上调,且有明显的剂量依赖性,并能被1 μg/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10 ng/ml SDF-1组比较,100 ng/ml SDF-1组有明显促进喉癌细胞增殖的作用(P0.05),并能被1 μg/ml的AMD3100阻断;③与对照组和10 ng/ml SDF-1组比较,100 ng/ml SDF-1组同样有明显的促喉癌细胞迁移和侵袭的作用(P0.05)
同样也能被1 μg/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 upregulated 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.
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