山东大学耳鼻喉眼学报 ›› 2015, Vol. 29 ›› Issue (3): 76-80.doi: 10.6040/j.issn.1673-3770.0.2015.154

• 论著 • 上一篇    下一篇

下调血管内皮细胞蛋白激酶CK2表达对血管增殖影响的体外研究

杜祥阁1, 张营春2, 颜昕2, 王翠2, 赵博军2   

  1. 1. 山东中医药大学第二临床学院, 山东 济南 250014;
    2. 山东大学附属省立医院眼科中心, 山东 济南 250021
  • 收稿日期:2015-04-13 修回日期:2015-05-13 发布日期:2015-06-16
  • 作者简介:杜祥阁。E-mail:duxiangge@126.com

In vitro study of the effect of Casein Kinase2 siRNA on anginogesis

DU Xiangge1, ZHANG Yingchun2, YAN Xin2, WANG Cui2, ZHAO Bojun2   

  1. 1. Second Clinical Medicine College, Shandong University of Traditional Chinese Medicine, Jinan 250014, Shandong, China;
    2. Department of Ophthalmology, Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China
  • Received:2015-04-13 Revised:2015-05-13 Published:2015-06-16
  • Contact: 赵博军。E-mail:zhaobojun@hotmail.com E-mail:zhaobojun@hotmail.com

摘要: 目的 探究蛋白激酶CK2表达下调对血管内皮细胞形成新生血管的能力以及对其细胞中血管内皮生长因子(VEGF)表达的影响。方法 化学合成siRNA转染人脐静脉内皮细胞(HUVECs), 用RT-PCR法和Western blotting法检测细胞转染效率;MTT法检测CK2表达下调对HUVECs细胞增殖的影响;成管实验和划痕实验检测CK2表达下调对细胞成管、迁移的影响;Real-time PCR和Western blotting法检测siRNA干扰蛋白激酶CK2表达后VEGF mRNA及蛋白的表达。结果 siRNA干扰蛋白激酶CK2达到了预期干扰率75%, 蛋白激酶CK2表达下调后抑制了HUVECs增殖、成管、迁移(P<0.05), 也抑制了VEGF mRNA及蛋白的表达。结论 蛋白激酶CK2表达下调可抑制VEGF的表达及HUVECs的增殖、成管、迁移。

关键词: 蛋白激酶CK2, siRNA, 血管内皮细胞, 视网膜新生血管, 血管内皮生长因子

Abstract: Objective To prove the effect of Casein Kinase2 siRNA on vascular endothelial growth factor(VEGF) expression and the angiogenic capacity in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were transfected with either Casein Kinase2 siRNA or negative control siRNA. Cell proliferation was analyzed by MTT assay; the angiogenic capacity was accessed by tube formation; cell migration was assessed by the wound-healing model. Gene and protein expressions of VEGF were examined by Real-time PCR and Western blotting methods. Results Casein Kinase2 siRNA decreased the expressions of Casein Kinase2 and declined HUVEC proliferation, tube formation and migration. It was also reduced the expression of VEGF mRNA and VEGF protein. Conclusion Our results demonstrated that Casein Kinase2 siRNA reduced the expression of VEGF and declined the capacity of angiogenesis in HUVECs.

Key words: Angiogenesis, Vascular endothelial growth factor, Casein Kinase 2, siRNA, Vascular endothelial cells

中图分类号: 

  • R774.1
[1] Campochiaro P A. Ocular neovascularization[J]. J Mol Med (Berl), 2013, 91(3):311-321.
[2] Dorrell M, Uusitalo-Jarvinen H, Aguilar E, et al. Ocular neovascularization: basic mechanisms and therapeutic advances[J]. Surv Ophthalmol, 2007, 52(1):3-19.
[3] Andrei A. Expression of protein kinase CK2 in astroglial cells of normal and neovascularized retina[J]. Am J Pathol, 2006, 168(5):1722-1736.
[4] Kramerov A A, Ljubimov A V. Focus on molecules:protein kinase CK2[J]. Exp Eye Res, 2012, 8(101):111-112.
[5] Barnstable C J, Tombran-Tink J. Neuroprotective and antiangiogenic actions of PEDF in the eye: molecular targets and therapeutic potential[J]. Prog Retin Eye Res, 2004, 23(5):561-577.
[6] Hou J, Lin L, Zhou W, et al. Identification of miRNomes in human liver and hepatocellular carcinoma reveals mir-199ab-3p as therapeutic target for hepatocellular carcinoma[J].Cancer Cell, 2011, 19(2):232-243.
[7] Ren Shancheng, Liu Yawei. Long noncoding RNA MALAT-1 is a new potential therapeutic target for castration resistant prostate cancer[J]. J Urol, 2013, 190(6):2278-2287.
[8] Duncan J S, Litchfield D W. Too much of a good thing: the role of protein kinase CK2 in tumorigenesis and prospects for therapeutic inhibition of CK2[J]. Biochim Biophys Acta, 2008, 1784(1):33-47.
[9] Dorrell M I, Aguilar E, Scheppke L, et al . Combination angiostatic therapy completely inhibits ocular and tumor angiogenesis[J]. Proc Natl Acad Sci USA, 2007, 104(3):967-972.
[10] Maloney S C, Godeiro K D, Odashiro A N, et al. Current and emerging concepts in the management of neovascular age-related macular degeneration[J]. Cardiovasc Hematol Agents Med Chem, 2007, 5(2):147-154.
[11] Ljubimov A V, Caballero S, Aoki A M, et al. Involvement of protein kinase CK2 in angiogenesis and retinal neovascularization[J]. Invest Ophthalmol Vis Sci, 2004, 45(12):4583-4591.
[12] Pagano M A, Poletto G, Di Maira G, et al. Tetrabromocinnamic acid (TBCA) and related compounds represent a new class of specific protein kinase CK2 inhibitors[J]. Chembiochem, 2007, 28(1):129-139.
[13] Golub A G, Yakovenko O Y, Prykhod'ko A O, et al. Evaluation of 4, 5, 6, 7-tetrahalogeno-1H-isoindole-1, 3(2H)-diones as inhibitors of human protein kinase CK2[J]. Biochim Biophys Acta, 2008, 1784(1):143-149.
[14] Zamore P D, Tuschl T, Sharp P A, et al. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals[J]. Cell, 2000, 101(1):25-33.
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