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Cytotechnology

, Volume 56, Issue 3, pp 219–231 | Cite as

Small interfering RNA knockdown of mini-TyrRS and mini-TrpRS effects angiogenesis in human umbilical vein endothelial cells in hypoxic culture

  • Rui Zeng
  • Yu-cheng Chen
  • Zhi ZengEmail author
  • Rui Liu
  • Ou Qiang
  • Xiao-fei Jiang
  • Xiao-xia Liu
  • Xian Li
  • Hao-yu Wang
Original Research

Abstract

Aim We studied the role of mini-TyrRS and mini-TrpRS in angiogenesis by using small interfering RNA-mediated mini-TyrRS/mini-TrpRS knockout in hypoxic culture of human umbilical vein endothelial cells. Methods SiRNA was used as the main method to inhibited the gene function. Silencing efficiency was assayed by real-time reverse transcription-polymerase chain reaction and western blotting. The angiogenic activity in vitro was evaluated by transwell migration assay and Matrigel-induced capillary tube formation in hypoxic culture. Cell proliferation was determined by crystal violet staining. Results The results showed that levels of the mini-TyrRS/mini-TrpRS gene and protein in mock transfection group and negative control group were higher, but noticeably decreased in experimental group. However, no significant difference was detected between mock transfection group and negative control group, but there was a statistically significant difference compared with experimental group. For mini-TyrRS-siRNA group, the cell migration, tube formation and the rate of cell proliferation were respectively inhibited by (47.4, 56.3, 65.4, 73.7%), (60.5, 69.1, 75.9, 83.6%) and (40.4, 56.2, 61.2, 68.0%). For mini-TrpRS-siRNA, were respectively increased by (18.0, 33.8, 45.1, 56.4%), (18.3, 31.2, 40.3, 45.7%) and (8.4, 26.4, 38.2, 46.6%). Conclusion These results indicated that angiogenesis is either stimulated by mini-TyrRS or inhibited by mini-TrpRS in matrigel models in hypoxic culture, raising the possibility that mini-TyrRS stimulates a common downstream signaling event. Thus, naturally occurring fragments of two proteins involved in translation, TyrRS and TrpRS, have opposing activity on endothelial cell angiogenesis in the matrigel assays. The opposing activities of the two tRNA synthetases suggest tight regulation of the balance between pro- and anti-angiogenic stimuli.

Keywords

Mini-TyrRS Mini-TrpRS Hypoxia Human umbilical vein endothelial cells siRNA Angiogenesis 

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Rui Zeng
    • 1
  • Yu-cheng Chen
    • 1
  • Zhi Zeng
    • 1
    Email author
  • Rui Liu
    • 2
  • Ou Qiang
    • 2
  • Xiao-fei Jiang
    • 1
  • Xiao-xia Liu
    • 3
  • Xian Li
    • 2
  • Hao-yu Wang
    • 1
  1. 1.Department of Cardiology, West China Hospital, School of Clinic MedicineSichuan UniversityChengduChina
  2. 2.Laboratory of Peptides Related with Human Diseases, The National Laboratory of BiomedicineSichuan UniversityChengduChina
  3. 3.Department of Epidemiology and Health Statistics, School of Public HealthSichuan UniversityChengduChina

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