缺氧環境下A549細胞促進人臍靜脈內皮細胞遷移及血管形成_《中國呼吸與危重監護雜志》

作者：

1. 中南大學湘雅醫院呼吸內科(湖南長沙 410008);;
2. 長沙市中心醫院呼吸內科(湖南長沙 410004);

關鍵詞：

低氧 A549細胞人臍靜脈內皮細胞低氧誘導因子1α 遷移偽足三維培養技術

DOI：

10.7507/1671-6205.2014043

視頻：

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目的研究在缺氧條件下A549細胞對人臍靜脈內皮細胞(HUVEC)遷移及微血管形成的影響。方法 A549細胞在常氧組(21%O₂)、缺氧組(2%O₂)、無氧組(0%O₂)中培養24 h后,通過倒置相差顯微鏡觀察細胞形態、MTT法檢測細胞增殖、免疫細胞化學技術檢測細胞內低氧誘導因子1α(HIF-1α)蛋白量以確定缺氧模型建立是否成功。隨后取常氧組、缺氧組的A549細胞上清液、HUVEC細胞培養液干預HUVEC,用細胞劃痕實驗、微絲綠色熒光染色方法觀察各組HUVEC的遷移情況及偽足形成,體外HUVEC三維培養技術觀察血管形成情況。結果與常氧組比較,缺氧組A549細胞的生長狀態好,增殖明顯;無氧組A549細胞生長狀態差,細胞數量減少;缺氧組及無氧組A549細胞均有HIF-1α表達,且缺氧組表達更加明顯。與細胞培養液干預組比較,缺氧上清液干預組和常氧上清液干預組A549細胞的HUVEC均有不同程度的遷移、偽足結構形成及血管管腔樣結構形成,且前者較后者明顯。結論缺氧環境下A549細胞生長狀態良好,增殖明顯,而無氧環境下則不利于A549細胞的生長。缺氧環境下A549細胞能促進HUVEC的遷移、偽足形成及血管形成。

引用本文： 胡贊, 李敏, 胡成平, 莫貴艷, 顧其華. 缺氧環境下A549細胞促進人臍靜脈內皮細胞遷移及血管形成. 中國呼吸與危重監護雜志, 2013, 12(2): 179-184. doi: 10.7507/1671-6205.2014043 復制

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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,陳永強.缺氧誘導因子-1與腫瘤細胞凋亡及血管生成關系的研究進展.中華腫瘤防治雜志,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,徐慶祥,李玉艷,等.新型喜樹堿類衍生物9-甲基哌嗪甲基-10-羥基喜樹堿氮氧化物對肺癌A549細胞缺氧誘導因子-1α表達的影響.中華實驗外科雜志,2012,29:1859-1860.
6. 王翎,馮春來,董寧征.老年原發性肺癌患者缺氧誘導因子-1α和葡萄糖轉運蛋白-1的表達及臨床意義.中華老年醫學雜志,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,劉濤,王偉,等.uPA、VEGF和微血管密度在卵巢惡性生殖細胞腫瘤中表達及其臨床意義研究.中華實用診斷與治療雜志,2011,25:548-551.
9. 顏浩,徐英.非小細胞肺癌中兩種雌激素受體亞型與血管內皮生長因子的表達及相關性研究.中國呼吸與危重監護雜志,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,張冠鑫,侯霄雷,等.沉默血管內皮生長因子及促血管生成素-2基因抑制裸鼠人肺腺癌移植瘤生長的實驗研究.中華臨床醫師雜志:電子版,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,蔣煒,王吉耀,等.轉化生長因子β1對肝星狀細胞遷移及細胞內Rho三磷酸鳥苷酶表達的影響.中華消化雜志,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.

《中國呼吸與危重監護雜志》

缺氧環境下A549細胞促進人臍靜脈內皮細胞遷移及血管形成

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《中國呼吸與危重監護雜志》

缺氧環境下A549細胞促進人臍靜脈內皮細胞遷移及血管形成

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