title: [Knockdown of RUNX3 inhibits hypoxia-induced endothelial-to-mesenchymal transition of human cardiac microvascular endothelial cells].
abstract: Objective To investigate the effects of Runt-related transcription factor 3 (RUNX3) knockdown on hypoxia-induced endothelial-to-mesenchymal transition (EndoMT) of human cardiac microvascular endothelial cells (HCMECs), and elucidate the underlying molecular mechanism. Methods HCMECs were cultured in hypoxic conditions and infected with RUNX3-RNAi lentivirus to knock-down the expression of RUNX3. Reverse transcription PCR was performed to detect the mRNA expressions of RUNX3 and EndoMT related genes such as CD31, vascular endothelial cadherin (VE-cadherin), α-smooth muscle actin (α-SMA) and fibroblast-specific protein-1 (FSP-1); Western blotting was used to determine the protein expressions of RUNX3, CD31, α-SMA and another molecules involved in EndoMT; and immunofluorescence cytochemistry was applied to observe the colocalization of CD31 and α-SMA. Results Hypoxia induced the transition of HCMECs to mesenchymal cells. Hypoxia up-regulated the expression of TGF-β2, Smad2/3, phosphorylation of Smad2/3 (p-Smad2/3), Notch-1, Hes1, and Hey1; knockdown of RUNX3 down-regulated the levels of Smad2/3, p-Smad2/3, Hes1, and Hey1 to different extents, and raised the levels of TGF-β2 and Notch-1. Conclusion Knockdown of RUNX3 in HCMECs attenuates hypoxia-induced EndoMT via partially inhibiting TGF-β and Notch signaling pathway.
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Nanchang University, Nanchang 330006; Department of Cardiology, Loudi Central Hospital, Loudi 417000, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Nanchang University, Nanchang 330006, China.*Corresponding author, E-mail: libingong08@163.com.
authors.forename: Bingong
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Cardiology, Loudi Central Hospital, Loudi 417000, China.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Surgical Oncology, Loudi Central Hospital, Loudi 417000, China.
authors.forename: Delong
authors.initials: D
authors.lastname: Wang
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Nanchang University, Nanchang 330006, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Zou
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Nanchang University, Nanchang 330006, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Ke
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Nanchang University, Nanchang 330006, China.
authors.forename: Yanqin
authors.initials: Y
authors.lastname: Hao
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D015703:Antigens, CD; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D015820:Cadherins; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D050677:Core Binding Factor Alpha 3 Subunit / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics / Q000502:physiology*; D006801:Humans; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071999:S100 Calcium-Binding Protein A4; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism; D000072056:Transcription Factor HES-1 / Q000235:genetics / Q000378:metabolism; D053781:Transforming Growth Factor beta2 / Q000235:genetics / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D000199:Actins; D015703:Antigens, CD; D051792:Basic Helix-Loop-Helix Proteins; D015820:Cadherins; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D050677:Core Binding Factor Alpha 3 Subunit; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D000071999:S100 Calcium-Binding Protein A4; D051899:Smad2 Protein; D051900:Smad3 Protein; D000072056:Transcription Factor HES-1; D053781:Transforming Growth Factor beta2; D000099152:Cadherin 5; C541116:ACTA2 protein, human; C485631:HEY1 protein, human; C493729:Runx3 protein, human; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; C077591:S100A4 protein, human; C078493:HES1 protein, human