A549 Cells Promote HUVEC Migration and Angiogenesis under Hypoxic Conditions_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

Department of Respiratory Medicine, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China;

Corresponding author：

Keywords：

Hypoxia; A549 cells; Human umbilical vein endothelial cells; Hypoxia-inducible factor-1α; Migration; Pseudopodia; Three-dimensional culture techniques

DOI：

10.7507/1671-6205.2014043

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Objective To observe the effects of A549 cells under hypoxicconditions on the migration of human umbilical vein endothelial cells (HUVECs) and microvascular formation. Methods After cultured for 24 h in normoxia condition(21% O₂),hypoxia condition (2% O₂),and anaerobic condition (0% O₂),respectively,morphology of A549 cells was observed with inverted phase contrast microscope,proliferation was detected by MTT assay,and intracellular hypoxia-inducible factor-1α (HIF-1α) protein was detected by immunocyto-chemical technique,for determining whether the hypoxia model is successful. Then A549 cells' supernatant in the normoxic group,the hypoxia group and HUVECs culture medium were taken to intervene HUVECs. The migration of HUVECs was observed with cell scratch test,pseudopodia formation of HUVECs was observed with microfilament green fluorescent staining method,and blood vessel formation was observed with three-dimensional culture techniques in vitro. Results Compared with the normoxic group,the growth of A549 cells was better in the hypoxia group with more proliferation,and was poor in the anaerobic group with decreased number of cells. A549 cells in the hypoxia group and the anaerobic group both expressed HIF-1α protein,which was more obvious in the anaerobic group. Compared with the HUVECs supernatant intervention group,the hypoxia supernatant intervention group and the normoxic supernatant intervention group both had varying degrees of migration,pseudopodia structure formation and vascular lumen sample structure formation,which were more obvious in the former group. Conclusion A549 cells in hypoxic environment grow very well,proliferated significantly,but anaerobic environment is not conducive to the growth of A549 cells which found to be apoptosis. A549 cells in hypoxic environment can promote HUVECs migration,pseudopodia formation and angiogenesis.

Citation： Hu Zan, Li Min, Hu Chengping, Mo Guiyan, Gu Qihua. A549 Cells Promote HUVEC Migration and Angiogenesis under Hypoxic Conditions. Chinese Journal of Respiratory and Critical Care Medicine, 2013, 12(2): 179-184. doi: 10.7507/1671-6205.2014043 Copy

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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.

Chinese Journal of Respiratory and Critical Care Medicine

A549 Cells Promote HUVEC Migration and Angiogenesis under Hypoxic Conditions

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