title: Effects of CASP5 gene overexpression on angiogenesis of HMEC-1 cells.
abstract: OBJECTIVE
  The efficacy of gene overexpression of CASP5, a caspase family member, in angiogenesis in vitro and its mechanisms were clarified.
  
  
  METHODS
  Human full-length CASP5 gene was delivered into human microvascular endothelial HMEC-1 cells by recombinant lentivirus. The infection was estimated by green fluorescent protein. MTT method was used to analyze the efficacy of gene overexpression in cell proliferation ability, and Matrigel was used to estimate its effects in angiogenesis ability of cells. Meanwhile, Western blot was used to analyze the effects of CASP5 gene overexpression on the expression levels of angpt-1, angpt-2, Tie2 and VEGF-1 in the cells, which were signaling pathway factors related to angiogenesis.
  
  
  RESULTS
  Recombinant lentivirus containing human full-length CASP5 gene was packed and purified successfully, with virus titer of 1×10(8) TU/ml. The recombinant lentivirus was used to infect HMEC-1 cells with MOI of 1, leading to a cell infection rate of 100%. There were no significant effects of CASP5 gene overexpression on both cell proliferation ability and the expression level of angpt-1. Meanwhile, expressions of angpt-2 and VEGF-1 were both enhanced, while Tie2 expression was inhibited. Results indicated that CASP5 gene overexpression promoted angiogenesis of HMEC-1 cells.
  
  
  CONCLUSIONS
  CASP5 gene overexpression significantly promoted angiogenesis ability of HMEC-1 cells, which was probably achieved by inhibiting angpt-1/Tie2 and promoting VEGF-1 signal pathway.
authors.affiliation: Department of Dermatology, Harbin Medical University Cancer Hospital Harbin 150040, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Dermatology, The Second Affiliated Hospital of Harbin Medical University Harbin 150010, China.
authors.forename: Yuzhen
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Dermatology, The First Affiliated Hospital of Harbin Medical University Harbin 150010, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Cai
authors.affiliation: Department of Dermatology, The Second Affiliated Hospital of Harbin Medical University Harbin 150010, China.
authors.forename: Bingxue
authors.initials: B
authors.lastname: Bai
authors.affiliation: Department of Dermatology, The First Affiliated Hospital of Harbin Medical University Harbin 150010, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Wang
mesh_terms: D042702:Angiopoietin-2 / Q000235:genetics / Q000378:metabolism; D020169:Caspases / Q000096:biosynthesis* / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000201:enzymology*; D004790:Enzyme Induction; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D018919:Neovascularization, Physiologic; D042787:Receptor, TIE-2 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
chemical_list: C586513:ANGPT2 protein, human; D042702:Angiopoietin-2; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042787:Receptor, TIE-2; C099217:CASP5 protein, human; D020169:Caspases
keywords: CASP5 gene; HMEC-1 cells; angiogenesis; caspase