Regulation of Angiopoietin-2 mRNA Levels in Bovine Microvascular Endothelial Cells by Cytokines and Hypoxia

From the Department of Morphology, University Medical Center, Geneva, Switzerland.

Abstract—Angiopoietin-2 (Ang2) is a ligand for the endothelial cell tyrosine kinase receptor Tie2 and counteracts blood vessel maturation/stability mediated by angiopoietin-1 (Ang1), the other known ligand of Tie2. Using degenerate oligonucleotides and reverse transcriptase–polymerase chain reaction, we have screened bovine microvascular endothelial (BME), aortic, lymphatic, pulmonary artery, and transformed fetal aortic endothelial cells, as well as rat smooth muscle cells for Ang1 and Ang2 expression. Except for high Ang2 mRNA levels found in BME cells, none of the endothelial cell types studied expressed appreciable levels of Ang1 or Ang2 mRNAs, whereas smooth muscle cells expressed both Ang1 and Ang2. BME cell Ang2 mRNA levels were increased by vascular endothelial growth factor (1.9- to 2.9-fold), basic fibroblast growth factor (1.6- to 2-fold), both cytokines in combination (2.9- to 4-fold), and hypoxia (3.1- to 5.6-fold) and were decreased by Ang1 (31% to 70%) or transforming growth factor-ß₁ (64% to 81%). Ang2 also decreased (60% to 82%) BME cell Ang2 mRNA. mRNA levels for the Tie1 or Tie2 receptors were only slightly modulated under the conditions described above. These findings suggest that the angiogenic effect of a number of regulators may be achieved in part through the regulation of an autocrine loop of Ang2 activity in microvascular endothelial cells.

Key Words: angiopoietin-2 • angiogenesis • hypoxia • cytokine • vascular remodeling

This article has been cited by other articles:

				P. Kumpers, J. Hellpap, S. David, R. Horn, H. Leitolf, H. Haller, and M. Haubitz Circulating angiopoietin-2 is a marker and potential mediator of endothelial cell detachment in ANCA-associated vasculitis with renal involvement Nephrol. Dial. Transplant., June 1, 2009; 24(6): 1845 - 1850. [Abstract] [Full Text] [PDF]

				E. M. Dioum, S. L. Clarke, K. Ding, J. J. Repa, and J. A. Garcia HIF-2{alpha}-Haploinsufficient Mice Have Blunted Retinal Neovascularization Due to Impaired Expression of a Proangiogenic Gene Battery Invest. Ophthalmol. Vis. Sci., June 1, 2008; 49(6): 2714 - 2720. [Abstract] [Full Text] [PDF]

				H. Ishimoto, K. Minegishi, T. Higuchi, M. Furuya, S. Asai, S. H. Kim, M. Tanaka, Y. Yoshimura, and R. B. Jaffe The Periphery of the Human Fetal Adrenal Gland Is a Site of Angiogenesis: Zonal Differential Expression and Regulation of Angiogenic Factors J. Clin. Endocrinol. Metab., June 1, 2008; 93(6): 2402 - 2408. [Abstract] [Full Text] [PDF]

				M. Mofarrahi, T. Nouh, S. Qureshi, L. Guillot, D. Mayaki, and S. N. A. Hussain Regulation of angiopoietin expression by bacterial lipopolysaccharide Am J Physiol Lung Cell Mol Physiol, May 1, 2008; 294(5): L955 - L963. [Abstract] [Full Text] [PDF]

				M. Szymczak, M. Murray, and N. Petrovic Modulation of angiogenesis by {omega}-3 polyunsaturated fatty acids is mediated by cyclooxygenases Blood, April 1, 2008; 111(7): 3514 - 3521. [Abstract] [Full Text] [PDF]

				L. Kugathasan, A. E. Dutly, Y. D. Zhao, Y. Deng, M. J. Robb, S. Keshavjee, and D. J. Stewart Role of Angiopoietin-1 in Experimental and Human Pulmonary Arterial Hypertension Chest, December 1, 2005; 128(6_suppl): 633S - 642S. [Abstract] [Full Text] [PDF]

				R. E. Nisato, G. Hosseini, C. Sirrenberg, G. S. Butler, T. Crabbe, A. J.P. Docherty, M. Wiesner, G. Murphy, C. M. Overall, S. L. Goodman, et al. Dissecting the Role of Matrix Metalloproteinases (MMP) and Integrin {alpha}v{beta}3 in Angiogenesis In vitro: Absence of Hemopexin C Domain Bioactivity, but Membrane-Type 1-MMP and {alpha}v{beta}3 Are Critical Cancer Res., October 15, 2005; 65(20): 9377 - 9387. [Abstract] [Full Text] [PDF]

				J S Mohan, P L Lip, A D Blann, D Bareford, and G Y H Lip The angiopoietin/Tie-2 system in proliferative sickle retinopathy: relation to vascular endothelial growth factor, its soluble receptor Flt-1 and von Willebrand factor, and to the effects of laser treatment Br J Ophthalmol, July 1, 2005; 89(7): 815 - 819. [Abstract] [Full Text] [PDF]

				J I Patel, P G Hykin, Z J Gregor, M Boulton, and I A Cree Angiopoietin concentrations in diabetic retinopathy Br J Ophthalmol, April 1, 2005; 89(4): 480 - 483. [Abstract] [Full Text] [PDF]

				A. Hegen, S. Koidl, K. Weindel, D. Marme, H. G. Augustin, and U. Fiedler Expression of Angiopoietin-2 in Endothelial Cells Is Controlled by Positive and Negative Regulatory Promoter Elements Arterioscler Thromb Vasc Biol, October 1, 2004; 24(10): 1803 - 1809. [Abstract] [Full Text] [PDF]

				R. Sandhu, K. Teichert-Kuliszewska, S. Nag, G. Proteau, M. J. Robb, A. I.M. Campbell, M. A. Kuliszewski, M. J.B. Kutryk, and D. J. Stewart Reciprocal regulation of angiopoietin-1 and angiopoietin-2 following myocardial infarction in the rat Cardiovasc Res, October 1, 2004; 64(1): 115 - 124. [Abstract] [Full Text] [PDF]

				R. E. Nisato, J. A. Harrison, R. Buser, L. Orci, C. Rinsch, R. Montesano, P. Dupraz, and M. S. Pepper Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span Am. J. Pathol., July 1, 2004; 165(1): 11 - 24. [Abstract] [Full Text] [PDF]

				U. Fiedler, M. Scharpfenecker, S. Koidl, A. Hegen, V. Grunow, J. M. Schmidt, W. Kriz, G. Thurston, and H. G. Augustin The Tie-2 ligand Angiopoietin-2 is stored in and rapidly released upon stimulation from endothelial cell Weibel-Palade bodies Blood, June 1, 2004; 103(11): 4150 - 4156. [Abstract] [Full Text] [PDF]

				N. Tsutsumi, Y. Yonemitsu, Y. Shikada, M. Onimaru, M. Tanii, S. Okano, K. Kaneko, M. Hasegawa, M. Hashizume, Y. Maehara, et al. Essential Role of PDGFR{alpha}-p70S6K Signaling in Mesenchymal Cells During Therapeutic and Tumor Angiogenesis In Vivo: Role of PDGFR{alpha} During Angiogenesis Circ. Res., May 14, 2004; 94(9): 1186 - 1194. [Abstract] [Full Text] [PDF]

				H.-P. Hammes, J. Lin, P. Wagner, Y. Feng, F. vom Hagen, T. Krzizok, O. Renner, G. Breier, M. Brownlee, and U. Deutsch Angiopoietin-2 Causes Pericyte Dropout in the Normal Retina: Evidence for Involvement in Diabetic Retinopathy Diabetes, April 1, 2004; 53(4): 1104 - 1110. [Abstract] [Full Text] [PDF]

				P. Pichiule, J. C. Chavez, and J. C. LaManna Hypoxic Regulation of Angiopoietin-2 Expression in Endothelial Cells J. Biol. Chem., March 26, 2004; 279(13): 12171 - 12180. [Abstract] [Full Text] [PDF]

				G. J. Hausman and R. L. Richardson Adipose tissue angiogenesis J Anim Sci, March 1, 2004; 82(3): 925 - 934. [Abstract] [Full Text] [PDF]

				A. Y. Chong, G. J. Caine, B. Freestone, A. D. Blann, and G. Y. H. Lip Plasma angiopoietin-1, angiopoietin-2, and angiopoietin receptor tie-2 levels in congestive heart failure J. Am. Coll. Cardiol., February 4, 2004; 43(3): 423 - 428. [Abstract] [Full Text] [PDF]

				T. VEIKKOLA, M. LOHELA, K. IKENBERG, T. MAKINEN, T. KORFF, A. SAARISTO, T. PETROVA, M. JELTSCH, H. G. AUGUSTIN, and K. ALITALO Intrinsic versus microenvironmental regulation of lymphatic endothelial cell phenotype and function FASEB J, November 1, 2003; 17(14): 2006 - 2013. [Abstract] [Full Text] [PDF]

				M. Yamakawa, L. X. Liu, T. Date, A. J. Belanger, K. A. Vincent, G. Y. Akita, T. Kuriyama, S. H. Cheng, R. J. Gregory, and C. Jiang Hypoxia-Inducible Factor-1 Mediates Activation of Cultured Vascular Endothelial Cells by Inducing Multiple Angiogenic Factors Circ. Res., October 3, 2003; 93(7): 664 - 673. [Abstract] [Full Text] [PDF]

				O. Feraud, C. Mallet, and I. Vilgrain Expressional Regulation of the Angiopoietin-1 and -2 and the Endothelial-Specific Receptor Tyrosine Kinase Tie2 in Adrenal Atrophy: A Study of Adrenocorticotropin-Induced Repair Endocrinology, October 1, 2003; 144(10): 4607 - 4615. [Abstract] [Full Text] [PDF]

				L. Zhang, N. Yang, J.-W. Park, D. Katsaros, S. Fracchioli, G. Cao, A. O'Brien-Jenkins, T. C. Randall, S. C. Rubin, and G. Coukos Tumor-derived Vascular Endothelial Growth Factor Up-Regulates Angiopoietin-2 in Host Endothelium and Destabilizes Host Vasculature, Supporting Angiogenesis in Ovarian Cancer Cancer Res., June 15, 2003; 63(12): 3403 - 3412. [Abstract] [Full Text] [PDF]

				Y. D. Zhao, A. I.M. Campbell, M. Robb, D. Ng, and D. J. Stewart Protective Role of Angiopoietin-1 in Experimental Pulmonary Hypertension Circ. Res., May 16, 2003; 92(9): 984 - 991. [Abstract] [Full Text] [PDF]

				S. Le Jan, C. Amy, A. Cazes, C. Monnot, N. Lamande, J. Favier, J. Philippe, M. Sibony, J.-M. Gasc, P. Corvol, et al. Angiopoietin-Like 4 Is a Proangiogenic Factor Produced during Ischemia and in Conventional Renal Cell Carcinoma Am. J. Pathol., May 1, 2003; 162(5): 1521 - 1528. [Abstract] [Full Text] [PDF]

				H. Takagi, S. Koyama, H. Seike, H. Oh, A. Otani, M. Matsumura, and Y. Honda Potential Role of the Angiopoietin/Tie2 System in Ischemia-Induced Retinal Neovascularization Invest. Ophthalmol. Vis. Sci., January 1, 2003; 44(1): 393 - 402. [Abstract] [Full Text] [PDF]

				A. Zakrzewicz, T. W. Secomb, and A. R. Pries Angioadaptation: Keeping the Vascular System in Shape Physiology, October 1, 2002; 17(5): 197 - 201. [Abstract] [Full Text] [PDF]

				J. Favier, P.-F. Plouin, P. Corvol, and J.-M. Gasc Angiogenesis and Vascular Architecture in Pheochromocytomas : Distinctive Traits in Malignant Tumors Am. J. Pathol., October 1, 2002; 161(4): 1235 - 1246. [Abstract] [Full Text] [PDF]

				E. Geva, D. G. Ginzinger, C. J. Zaloudek, D. H. Moore, A. Byrne, and R. B. Jaffe Human Placental Vascular Development: Vasculogenic and Angiogenic (Branching and Nonbranching) Transformation Is Regulated by Vascular Endothelial Growth Factor-A, Angiopoietin-1, and Angiopoietin-2 J. Clin. Endocrinol. Metab., September 1, 2002; 87(9): 4213 - 4224. [Abstract] [Full Text] [PDF]

				G.C. Weston, I. Haviv, and P.A.W. Rogers Microarray analysis of VEGF-responsive genes in myometrial endothelial cells Mol. Hum. Reprod., September 1, 2002; 8(9): 855 - 863. [Abstract] [Full Text] [PDF]

				P. Hewett, S. Nijjar, M. Shams, S. Morgan, J. Gupta, and A. Ahmed Down-Regulation of Angiopoietin-1 Expression in Menorrhagia Am. J. Pathol., March 1, 2002; 160(3): 773 - 780. [Abstract] [Full Text] [PDF]

				J. Li, N. W. Shworak, and M. Simons Increased responsiveness of hypoxic endothelial cells to FGF2 is mediated by HIF-1{alpha}-dependent regulation of enzymes involved in synthesis of heparan sulfate FGF2-binding sites J. Cell Sci., January 5, 2002; 115(9): 1951 - 1959. [Abstract] [Full Text] [PDF]

				T. MAKINEN and K. ALITALO Molecular Mechanisms of Lymphangiogenesis Cold Spring Harb Symp Quant Biol, January 1, 2002; 67(0): 189 - 196. [Abstract] [PDF]

				Y. Yu, J. Varughese, L. F. Brown, J. B. Mulliken, and J. Bischoff Increased Tie2 Expression, Enhanced Response to Angiopoietin-1, and Dysregulated Angiopoietin-2 Expression in Hemangioma-Derived Endothelial Cells Am. J. Pathol., December 1, 2001; 159(6): 2271 - 2280. [Abstract] [Full Text] [PDF]

				K. Abdulmalek, F. Ashur, N. Ezer, F. Ye, S. Magder, and S. N. A. Hussain Differential expression of Tie-2 receptors and angiopoietins in response to in vivo hypoxia in rats Am J Physiol Lung Cell Mol Physiol, September 1, 2001; 281(3): L582 - L590. [Abstract] [Full Text] [PDF]

				K. Koga, T. Todaka, M. Morioka, J.-i. Hamada, Y. Kai, S. Yano, A. Okamura, N. Takakura, T. Suda, and Y. Ushio Expression of Angiopoietin-2 in Human Glioma Cells and Its Role for Angiogenesis Cancer Res., August 1, 2001; 61(16): 6248 - 6254. [Abstract] [Full Text] [PDF]

				J. D. Ramsden, H. C. Cocks, M. Shams, S. Nijjar, J. C. Watkinson, M. C. Sheppard, A. Ahmed, and M. C. Eggo Tie-2 Is Expressed on Thyroid Follicular Cells, Is Increased in Goiter, and Is Regulated by Thyrotropin through Cyclic Adenosine 3',5'-Monophosphate J. Clin. Endocrinol. Metab., June 1, 2001; 86(6): 2709 - 2716. [Abstract] [Full Text] [PDF]

				E. Audero, I. Cascone, I. Zanon, S. C. Previtali, R. Piva, D. Schiffer, and F. Bussolino Expression of Angiopoietin-1 in Human Glioblastomas Regulates Tumor-Induced Angiogenesis : In Vivo and In Vitro Studies Arterioscler Thromb Vasc Biol, April 1, 2001; 21(4): 536 - 541. [Abstract] [Full Text] [PDF]

				A. Otani, H. Takagi, H. Oh, S. Koyama, and Y. Honda Angiotensin II Induces Expression of the Tie2 Receptor Ligand, Angiopoietin-2, in Bovine Retinal Endothelial Cells Diabetes, April 1, 2001; 50(4): 867 - 875. [Abstract] [Full Text]

				T. Etoh, H. Inoue, S. Tanaka, G. F. Barnard, S. Kitano, and M. Mori Angiopoietin-2 Is Related to Tumor Angiogenesis in Gastric Carcinoma: Possible in Vivo Regulation via Induction of Proteases Cancer Res., March 1, 2001; 61(5): 2145 - 2153. [Abstract] [Full Text]

				I. Kim, S.-O. Moon, C.-Y. Han, Y. K. Pak, S. K. Moon, J. J. Kim, and G. Y. Koh The angiopoietin-tie2 system in coronary artery endothelium prevents oxidized low-density lipoprotein-induced apoptosis Cardiovasc Res, March 1, 2001; 49(4): 872 - 881. [Abstract] [Full Text] [PDF]

				K. Teichert-Kuliszewska, P. C. Maisonpierre, N. Jones, A. I.M. Campbell, Z. Master, M. P. Bendeck, K. Alitalo, D. J. Dumont, G. D. Yancopoulos, and D. J. Stewart Biological action of angiopoietin-2 in a fibrin matrix model of angiogenesis is associated with activation of Tie2 Cardiovasc Res, February 16, 2001; 49(3): 659 - 670. [Abstract] [Full Text] [PDF]

				N. C. Denko and A. J. Giaccia Tumor Hypoxia, the Physiological Link between Trousseau's Syndrome (Carcinoma-induced Coagulopathy) and Metastasis Cancer Res., February 1, 2001; 61(3): 795 - 798. [Full Text]

				T. KORFF, S. KIMMINA, G. MARTINY-BARON, and H. G. AUGUSTIN Blood vessel maturation in a 3-dimensional spheroidal coculture model: direct contact with smooth muscle cells regulates endothelial cell quiescence and abrogates VEGF responsiveness FASEB J, February 1, 2001; 15(2): 447 - 457. [Abstract] [Full Text] [PDF]

				S. Fujiyama, H. Matsubara, Y. Nozawa, K. Maruyama, Y. Mori, Y. Tsutsumi, H. Masaki, Y. Uchiyama, Y. Koyama, A. Nose, et al. Angiotensin AT1 and AT2 Receptors Differentially Regulate Angiopoietin-2 and Vascular Endothelial Growth Factor Expression and Angiogenesis by Modulating Heparin Binding-Epidermal Growth Factor (EGF)-Mediated EGF Receptor Transactivation Circ. Res., January 19, 2001; 88(1): 22 - 29. [Abstract] [Full Text] [PDF]

				S. E. Bell, A. Mavila, R. Salazar, K. J. Bayless, S. Kanagala, S. A. Maxwell, and G. E. Davis Differential gene expression during capillary morphogenesis in 3D collagen matrices: regulated expression of genes involved in basement membrane matrix assembly, cell cycle progression, cellular differentiation and G-protein signaling J. Cell Sci., January 8, 2001; 114(15): 2755 - 2773. [Abstract] [Full Text] [PDF]

				H. Ding, L. Roncari, X. Wu, N. Lau, P. Shannon, A. Nagy, and A. Guha Expression and hypoxic regulation of angiopoietins in human astrocytomas Neuro-oncol, January 1, 2001; 3(1): 1 - 10. [Abstract] [PDF]

				T.M. Hazzard, L.K. Christenson, and R.L. Stouffer Changes in expression of vascular endothelial growth factor and angiopoietin-1 and -2 in the macaque corpus luteum during the menstrual cycle Mol. Hum. Reprod., November 1, 2000; 6(11): 993 - 998. [Abstract] [Full Text] [PDF]

				H. Beck, T. Acker, C. Wiessner, P. R. Allegrini, and K. H. Plate Expression of Angiopoietin-1, Angiopoietin-2, and Tie Receptors after Middle Cerebral Artery Occlusion in the Rat Am. J. Pathol., November 1, 2000; 157(5): 1473 - 1483. [Abstract] [Full Text] [PDF]

				C. Willam, P. Koehne, J. S. Jurgensen, M. Grafe, K. D. Wagner, S. Bachmann, U. Frei, and K.-U. Eckardt Tie2 Receptor Expression Is Stimulated by Hypoxia and Proinflammatory Cytokines in Human Endothelial Cells Circ. Res., September 1, 2000; 87(5): 370 - 377. [Abstract] [Full Text] [PDF]

				M. Bongrazio, C. Baumann, A. Zakrzewicz, A. R Pries, and P. Gaehtgens Evidence for modulation of genes involved in vascular adaptation by prolonged exposure of endothelial cells to shear stress Cardiovasc Res, August 1, 2000; 47(2): 384 - 393. [Abstract] [Full Text] [PDF]

				S. J. Mandriota, C. Pyke, C. Di Sanza, P. Quinodoz, B. Pittet, and M. S. Pepper Hypoxia-Inducible Angiopoietin-2 Expression Is Mimicked by Iodonium Compounds and Occurs in the Rat Brain and Skin in Response to Systemic Hypoxia and Tissue Ischemia Am. J. Pathol., June 1, 2000; 156(6): 2077 - 2089. [Abstract] [Full Text] [PDF]

				T. Veikkola, M. Karkkainen, L. Claesson-Welsh, and K. Alitalo Regulation of Angiogenesis via Vascular Endothelial Growth Factor Receptors Cancer Res., January 1, 2000; 60(2): 203 - 212. [Full Text]

				J. Lauren, Y. Gunji, and K. Alitalo Is Angiopoietin-2 Necessary for the Initiation of Tumor Angiogenesis? Am. J. Pathol., November 1, 1998; 153(5): 1333 - 1339. [Full Text] [PDF]

				B. Cohen, D. Barkan, Y. Levy, I. Goldberg, E. Fridman, J. Kopolovic, and M. Rubinstein Leptin Induces Angiopoietin-2 Expression in Adipose Tissues J. Biol. Chem., March 9, 2001; 276(11): 7697 - 7700. [Abstract] [Full Text] [PDF]

				Y. Xu and Q. Yu Angiopoietin-1, Unlike Angiopoietin-2, Is Incorporated into the Extracellular Matrix via Its Linker Peptide Region J. Biol. Chem., September 7, 2001; 276(37): 34990 - 34998. [Abstract] [Full Text] [PDF]

				I. Kim, J.-H. Kim, Y. S. Ryu, S. H. Jung, J. J. Nah, and G. Y. Koh Characterization and Expression of a Novel Alternatively Spliced Human Angiopoietin-2 J. Biol. Chem., June 9, 2000; 275(24): 18550 - 18556. [Abstract] [Full Text] [PDF]