title: Novel role for p56/Lck in regulation of endothelial cell survival and angiogenesis.
abstract: Previous studies have demonstrated that cleaved high-molecular-weight kininogen (HKa) induces endothelial apoptosis and inhibits angiogenesis and have suggested that this occurs through inhibition of Src family kinases. This study assessed the role of tyrosine-protein kinase Lck (p56/Lck) in this pathway. We analyzed early events leading to apoptosis of human endothelial cells exposed to HKa. The role of p56/Lck was investigated using short interfering (si) RNA knockdown and lentivirus expression in assays of endothelial tube formation, sprouting of neovessels from murine aorta, and angiogenesis in Matrigel plugs. HKa stimulated expression and phosphorylation of p56/Lck. siRNA knockdown of p56/Lck promoted endothelial proliferation and blocked HKa-induced apoptosis and activation of p53, Bax, and Bak. Lentivirus expression of p56/Lck in endothelial cells induced apoptosis and blocked tube formation. Expression of p56/Lck in murine aortic rings blocked sprouting angiogenesis. Lentivirus expressing p56/Lck blocked angiogenesis in Matrigel plugs, while p56/Lck short hairpin RNA inhibited the antiangiogenic effect of HKa. Scrambled siRNAs and empty lentiviral vectors were used in all experiments. Apoptosis of proliferating endothelial cells and inhibition of angiogenesis by HKa requires p56/Lck. This suggests a novel role for p56/Lck in regulation of endothelial cell survival and angiogenesis.-Betapudi, V., Shukla, M., Alluri, R., Merkulov, S., McCrae, K. R. Novel role for p56/Lck in regulation of endothelial cell survival and angiogenesis.
authors.affiliation: Department of Cellular and Molecular Medicine Cleveland Clinic, Cleveland, Ohio, USA; Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio, USA; and.
authors.forename: Venkaiah
authors.initials: V
authors.lastname: Betapudi
authors.affiliation: Department of Cellular and Molecular Medicine Cleveland Clinic, Cleveland, Ohio, USA.
authors.forename: Meenal
authors.initials: M
authors.lastname: Shukla
authors.affiliation: Department of Cellular and Molecular Medicine Cleveland Clinic, Cleveland, Ohio, USA.
authors.forename: Ravi
authors.initials: R
authors.lastname: Alluri
authors.affiliation: Department of Cellular and Molecular Medicine Cleveland Clinic, Cleveland, Ohio, USA.
authors.forename: Sergei
authors.initials: S
authors.lastname: Merkulov
authors.affiliation: Department of Cellular and Molecular Medicine Cleveland Clinic, Cleveland, Ohio, USA; Taussig Cancer Institute, Cleveland Clinic, Cleveland, Ohio, USA mccraek@ccf.org.
authors.forename: Keith R
authors.initials: KR
authors.lastname: McCrae
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D002470:Cell Survival; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000235:genetics*; D018919:Neovascularization, Physiologic; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D043925:Angiogenesis Inducing Agents; D034741:RNA, Small Interfering; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
keywords: apoptosis; kininogen; p53; tyrosine protein kinase Lck