title: Silencing of Transient Receptor Potential Channel 4 Alleviates oxLDL-induced Angiogenesis in Human Coronary Artery Endothelial Cells by Inhibition of VEGF and NF-κB.
abstract: BACKGROUND
  Transient receptor potential channel 4 (TRPC4) plays central roles in endothelial cell function. The aim of this study was to investigate the silencing effects of TRPC4 on oxidized low-density lipoprotein (oxLDL)-induced angiogenesis in human coronary artery endothelial cells (HCAECs), as well as the underlying molecular mechanism involved in this process.
  
  
  METHODS
  HCAECs were transfected with small interfering RNA (siRNA) targeting TRPC4 (TRPC4-siRNA) or with a negative control (NC)-siRNA. The expression of TRPC4 was confirmed by real-time polymerase chain reaction (RT-PCR) and Western blotting. After the siRNA transfection, oxLDL was added to the medium. Cell proliferation, migration, and in vitro angiogenesis were determined by bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA), Transwell assay and scratch-wound assay, respectively, and tube formation on Matrigel. Expression of vascular endothelial growth factor (VEGF) and nuclear factor (NF)-κB p65 were assessed by Western blotting.
  
  
  RESULTS
  Both the mRNA and protein levels of TRPC4 were significantly reduced by transfection with TRPC4-siRNA compared to the control group or NC-siRNA group (P<0.05). Silencing of TRPC4 significantly decreased the cell proliferation, migration, and tube formation (all P<0.05). Furthermore, the expression levels of VEGF and NF-κB p65 were markedly lowered by silencing of TRPC4 in HCAECs.
  
  
  CONCLUSIONS
  These results suggest that silencing of TRPC4 alleviates angiogenesis induced by oxLDL in HCAECs through inactivation of VEGF and NF-κB. Suppression of TRPC4 might be an alternative therapeutic strategy for atherosclerotic neovascularization.
authors.affiliation: Department of Pathology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
authors.forename: Wen
authors.initials: W
authors.lastname: Qin
authors.affiliation: Department of Reproductive Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
authors.forename: Wei
authors.initials: W
authors.lastname: Xie
authors.affiliation: Department of Endocrinology and Metabolism, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China (mainland).
authors.forename: Ning
authors.initials: N
authors.lastname: Xia
authors.affiliation: , Guangxi Medical University, Nanning, Guangxi, China (mainland).
authors.forename: Qinglin
authors.initials: Q
authors.lastname: He
authors.affiliation: , Guangxi Medical University, Nanning, Guangxi, China (mainland).
authors.forename: Tianwei
authors.initials: T
authors.lastname: Sun
mesh_terms: D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003331:Coronary Vessels / Q000166:cytology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D020868:Gene Silencing / Q000187:drug effects*; D006801:Humans; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D050052:TRPC Cation Channels / Q000378:metabolism*; D051996:Transcription Factor RelA / Q000378:metabolism*; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D008077:Lipoproteins, LDL; D050052:TRPC Cation Channels; C489571:TRPC4 ion channel; D051996:Transcription Factor RelA; D042461:Vascular Endothelial Growth Factor A; C099252:oxidized low density lipoprotein