缺氧环境下A549细胞促进人脐静脉内皮细胞迁移及血管形成_《中国呼吸与危重监护杂志》

作者：

1. 中南大学湘雅医院呼吸内科(湖南长沙 410008);;
2. 长沙市中心医院呼吸内科(湖南长沙 410004);

关键词：

低氧 A549细胞人脐静脉内皮细胞低氧诱导因子1α 迁移伪足三维培养技术

DOI：

10.7507/1671-6205.2014043

视频：

导出 下载 收藏 扫码 引用

摘要 全文 图表 视频 参考文献 施引文献 补充材料

目的研究在缺氧条件下A549细胞对人脐静脉内皮细胞(HUVEC)迁移及微血管形成的影响。方法 A549细胞在常氧组(21%O₂)、缺氧组(2%O₂)、无氧组(0%O₂)中培养24 h后,通过倒置相差显微镜观察细胞形态、MTT法检测细胞增殖、免疫细胞化学技术检测细胞内低氧诱导因子1α(HIF-1α)蛋白量以确定缺氧模型建立是否成功。随后取常氧组、缺氧组的A549细胞上清液、HUVEC细胞培养液干预HUVEC,用细胞划痕实验、微丝绿色荧光染色方法观察各组HUVEC的迁移情况及伪足形成,体外HUVEC三维培养技术观察血管形成情况。结果与常氧组比较,缺氧组A549细胞的生长状态好,增殖明显;无氧组A549细胞生长状态差,细胞数量减少;缺氧组及无氧组A549细胞均有HIF-1α表达,且缺氧组表达更加明显。与细胞培养液干预组比较,缺氧上清液干预组和常氧上清液干预组A549细胞的HUVEC均有不同程度的迁移、伪足结构形成及血管管腔样结构形成,且前者较后者明显。结论缺氧环境下A549细胞生长状态良好,增殖明显,而无氧环境下则不利于A549细胞的生长。缺氧环境下A549细胞能促进HUVEC的迁移、伪足形成及血管形成。

引用本文： 胡赞, 李敏, 胡成平, 莫贵艳, 顾其华. 缺氧环境下A549细胞促进人脐静脉内皮细胞迁移及血管形成. 中国呼吸与危重监护杂志, 2013, 12(2): 179-184. doi: 10.7507/1671-6205.2014043 复制

1.	Ausprunk DH,Folkman J.Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis.Microvasc Res,1977,14:53-65.
2.	Bergers G,Benjami LE.Tumorigenesis and the angiogenie switch.Nat Rev Cancer,2003,3:401-410.
3.	李玉梅,陈永强.缺氧诱导因子-1与肿瘤细胞凋亡及血管生成关系的研究进展.中华肿瘤防治杂志,2008,15:948-950.
4.	Lee JW,Bae SH,Jeong JW,et al.Hypoxia-inducible factor (HIF-1)alpha: its protein stability and biological functions.Exp Mol Med,2004,36:1-12.
5.	徐益文,徐庆祥,李玉艳,等.新型喜树碱类衍生物9-甲基哌嗪甲基-10-羟基喜树碱氮氧化物对肺癌A549细胞缺氧诱导因子-1α表达的影响.中华实验外科杂志,2012,29:1859-1860.
6.	王翎,冯春来,董宁征.老年原发性肺癌患者缺氧诱导因子-1α和葡萄糖转运蛋白-1的表达及临床意义.中华老年医学杂志,2009,28:128-132.
7.	Li QF,Wang XR,Yang YW,et al.Hypoxia upregulates hypoxia inducible factor(HIF)-3 alpha expression in lung epithelial cells:characterization and comparison with HIF-1alpha.Cell Res,2006,16:548-558.
8.	王靖雯,刘涛,王伟,等.uPA、VEGF和微血管密度在卵巢恶性生殖细胞肿瘤中表达及其临床意义研究.中华实用诊断与治疗杂志,2011,25:548-551.
9.	颜浩,徐英.非小细胞肺癌中两种雌激素受体亚型与血管内皮生长因子的表达及相关性研究.中国呼吸与危重监护杂志,2010,9:287-290.
10.	McMullen M,Keller R,Sussman M,et al.Vascular endothelial growth factor-mediated activation of p38 is dependent upon Src and RAFTK/Pyk2.Oncogene,2004,23:1275-1282.
11.	Yoshizuka N,Chen RM,Xu Z,et al.A novel function of p38-regulated/activated kinase in endothelial cell migration and tumor angiogenesis.Mol Cell Biol,2012,32:606-618.
12.	李白翎,张冠鑫,侯霄雷,等.沉默血管内皮生长因子及促血管生成素-2基因抑制裸鼠人肺腺癌移植瘤生长的实验研究.中华临床医师杂志:电子版,2011,5:3756-3762.
13.	El-Sibai M,Nalbant P,Pang H,et al.Cdc42 is required for EGF-stimulated protrusion and motility in MTLn3 carcinoma cells.J Cell Sci,2007,120:3465-3474.
14.	李蕾,蒋炜,王吉耀,等.转化生长因子β1对肝星状细胞迁移及细胞内Rho三磷酸鸟苷酶表达的影响.中华消化杂志,2006,8:527-530.
15.	Laurent L,Le BF,Huot J.Endothelial Cell Migration During Angiogene.Circ Res,2007,100:782-794.
16.	Vader P,van der Meel R,Symons MH,et al.Examining the role of Rac1 in tumor angiogenesis and growth:a clinically relevant RNAi-mediated approach.Angiogenesis,2011,14:457-466.
17.	Wang W,Wu F,Fang F,et al.RhoC is essential for angiogenesis induced by hepatocellular carcinoma cells via regulation of endothelial cell organization.Cancer Sci,2008,99:2012-2018.
18.	Lamalice L,Houle F,Jourdan G,et al.Phosphorylation of tyrosine 1214 on VEGFR2 is required for VEGF-induced activation of Cdc42 upstream of SAPK2/p38.Oncogene,2004,23:434-445.
19.	Kitamura T,Asai N,Enomoto A,et al.Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin.Nat Cell Biol,2008,10:329-337.
20.	Franzdóttir SR,Axelsson IT,Arason AJ,et al.Airway branching morphogenesis in three dimensional culture.Respir Res,2010,11:162.
21.	Liu Y,Markov DA,Wikswo JP,et al.Microfabricated scaffold-guided endothelial morphogenesis in three-dimensional culture.Biomed Microdevices,2011,13:837-846.
22.	Kitel R,CzarneckaJ,Rusin A.Three-dimensional cell.cultures.Applications in basic science and biotechnology.Postepy Biochem,2013,59:305-314.
23.	Arya N,Sardana V,Saxena M,et al.Recapitulating tumour microenvironment in chitosan-gelatin three-dimensional scaffolds:an improved in vitro tumour model.J R Soc Interface,2012,9:3288-3302.
24.	Moreno-Bueno G,Peinado H,Molina P,et al.The morphological and molecular features of the epithelial-to-mesenchymal transition.Nat Protoc,2009,4:1591-1613.

1. Ausprunk DH,Folkman J.Migration and proliferation of endothelial cells in preformed and newly formed blood vessels during tumor angiogenesis.Microvasc Res,1977,14:53-65.
2. Bergers G,Benjami LE.Tumorigenesis and the angiogenie switch.Nat Rev Cancer,2003,3:401-410.
3. 李玉梅,陈永强.缺氧诱导因子-1与肿瘤细胞凋亡及血管生成关系的研究进展.中华肿瘤防治杂志,2008,15:948-950.
4. Lee JW,Bae SH,Jeong JW,et al.Hypoxia-inducible factor (HIF-1)alpha: its protein stability and biological functions.Exp Mol Med,2004,36:1-12.
5. 徐益文,徐庆祥,李玉艳,等.新型喜树碱类衍生物9-甲基哌嗪甲基-10-羟基喜树碱氮氧化物对肺癌A549细胞缺氧诱导因子-1α表达的影响.中华实验外科杂志,2012,29:1859-1860.
6. 王翎,冯春来,董宁征.老年原发性肺癌患者缺氧诱导因子-1α和葡萄糖转运蛋白-1的表达及临床意义.中华老年医学杂志,2009,28:128-132.
7. Li QF,Wang XR,Yang YW,et al.Hypoxia upregulates hypoxia inducible factor(HIF)-3 alpha expression in lung epithelial cells:characterization and comparison with HIF-1alpha.Cell Res,2006,16:548-558.
8. 王靖雯,刘涛,王伟,等.uPA、VEGF和微血管密度在卵巢恶性生殖细胞肿瘤中表达及其临床意义研究.中华实用诊断与治疗杂志,2011,25:548-551.
9. 颜浩,徐英.非小细胞肺癌中两种雌激素受体亚型与血管内皮生长因子的表达及相关性研究.中国呼吸与危重监护杂志,2010,9:287-290.
10. McMullen M,Keller R,Sussman M,et al.Vascular endothelial growth factor-mediated activation of p38 is dependent upon Src and RAFTK/Pyk2.Oncogene,2004,23:1275-1282.
11. Yoshizuka N,Chen RM,Xu Z,et al.A novel function of p38-regulated/activated kinase in endothelial cell migration and tumor angiogenesis.Mol Cell Biol,2012,32:606-618.
12. 李白翎,张冠鑫,侯霄雷,等.沉默血管内皮生长因子及促血管生成素-2基因抑制裸鼠人肺腺癌移植瘤生长的实验研究.中华临床医师杂志:电子版,2011,5:3756-3762.
13. El-Sibai M,Nalbant P,Pang H,et al.Cdc42 is required for EGF-stimulated protrusion and motility in MTLn3 carcinoma cells.J Cell Sci,2007,120:3465-3474.
14. 李蕾,蒋炜,王吉耀,等.转化生长因子β1对肝星状细胞迁移及细胞内Rho三磷酸鸟苷酶表达的影响.中华消化杂志,2006,8:527-530.
15. Laurent L,Le BF,Huot J.Endothelial Cell Migration During Angiogene.Circ Res,2007,100:782-794.
16. Vader P,van der Meel R,Symons MH,et al.Examining the role of Rac1 in tumor angiogenesis and growth:a clinically relevant RNAi-mediated approach.Angiogenesis,2011,14:457-466.
17. Wang W,Wu F,Fang F,et al.RhoC is essential for angiogenesis induced by hepatocellular carcinoma cells via regulation of endothelial cell organization.Cancer Sci,2008,99:2012-2018.
18. Lamalice L,Houle F,Jourdan G,et al.Phosphorylation of tyrosine 1214 on VEGFR2 is required for VEGF-induced activation of Cdc42 upstream of SAPK2/p38.Oncogene,2004,23:434-445.
19. Kitamura T,Asai N,Enomoto A,et al.Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin.Nat Cell Biol,2008,10:329-337.
20. Franzdóttir SR,Axelsson IT,Arason AJ,et al.Airway branching morphogenesis in three dimensional culture.Respir Res,2010,11:162.
21. Liu Y,Markov DA,Wikswo JP,et al.Microfabricated scaffold-guided endothelial morphogenesis in three-dimensional culture.Biomed Microdevices,2011,13:837-846.
22. Kitel R,CzarneckaJ,Rusin A.Three-dimensional cell.cultures.Applications in basic science and biotechnology.Postepy Biochem,2013,59:305-314.
23. Arya N,Sardana V,Saxena M,et al.Recapitulating tumour microenvironment in chitosan-gelatin three-dimensional scaffolds:an improved in vitro tumour model.J R Soc Interface,2012,9:3288-3302.
24. Moreno-Bueno G,Peinado H,Molina P,et al.The morphological and molecular features of the epithelial-to-mesenchymal transition.Nat Protoc,2009,4:1591-1613.

《中国呼吸与危重监护杂志》

缺氧环境下A549细胞促进人脐静脉内皮细胞迁移及血管形成

摘要 全文 图表 视频 参考文献 施引文献 补充材料

上一篇

下一篇

Format

Content

《中国呼吸与危重监护杂志》

缺氧环境下A549细胞促进人脐静脉内皮细胞迁移及血管形成

摘要 全文 图表 视频 参考文献 施引文献 补充材料

上一篇

下一篇

Format

Content

摘要全文图表视频参考文献施引文献补充材料