OBJECTIVE: In this study, the effects of survivin (SVV) on angiogenesis were evaluated in vitro and in vivo. METHODS: The adenovirus (Ad)-mediated murine SVV gene was transfected into rat aortic endothelial cells (RAECs). RAECs expressing green fluorescent protein after transfection with Ad served as a negative control and those without transfection as a blank control. Then, the SVV mRNA was detected by quantitative real time RT-PCR. The SVV protein, cell cycle and apoptosis related proteins, and matrix metalloproteinase (MMPs) were detected by western blot assay. Immunofluorescence staining was conducted for proliferating cell nuclear antigen and MTT assay for cell viability. Transwell and matrigel chamber assay were employed to assess the migration and invasion of cells after transfection. TUNEL staining and flow cytometry were performed to detect the apoptotic REACs after treatment with anti-Fas antibody. Tube formation in matrigel membranes and matrigel plugs assay in nude mice were employed to confirm the angiogenic capacity in vitro and in vivo, respectively. RESULTS: The mRNA and protein expressions of SVV increased significantly in SVV transfected cells. The SVV transfected cells showed increased cell proliferation, up-regulated expressions of cell cycle proteins, enhanced invasiveness and migration activities and increased expressions of MMP-2, 7 and 9. In addition, SVV protected against apoptosis of RAECs by inactivating caspase-3, 8 and 9. The tube formation and matrigel plugs assays showed SVV significantly increased blood vessels in vitro and in vivo. CONCLUSION: SVV may act as an angiogenic factor and used for therapeutic angiogenesis in peripheral arterial diseases.
OBJECTIVE: In this study, the effects of survivin (SVV) on angiogenesis were evaluated in vitro and in vivo. METHODS: The adenovirus (Ad)-mediated murineSVV gene was transfected into rat aortic endothelial cells (RAECs). RAECs expressing green fluorescent protein after transfection with Ad served as a negative control and those without transfection as a blank control. Then, the SVV mRNA was detected by quantitative real time RT-PCR. The SVV protein, cell cycle and apoptosis related proteins, and matrix metalloproteinase (MMPs) were detected by western blot assay. Immunofluorescence staining was conducted for proliferating cell nuclear antigen and MTT assay for cell viability. Transwell and matrigel chamber assay were employed to assess the migration and invasion of cells after transfection. TUNEL staining and flow cytometry were performed to detect the apoptotic REACs after treatment with anti-Fas antibody. Tube formation in matrigel membranes and matrigel plugs assay in nude mice were employed to confirm the angiogenic capacity in vitro and in vivo, respectively. RESULTS: The mRNA and protein expressions of SVV increased significantly in SVV transfected cells. The SVV transfected cells showed increased cell proliferation, up-regulated expressions of cell cycle proteins, enhanced invasiveness and migration activities and increased expressions of MMP-2, 7 and 9. In addition, SVV protected against apoptosis of RAECs by inactivating caspase-3, 8 and 9. The tube formation and matrigel plugs assays showed SVV significantly increased blood vessels in vitro and in vivo. CONCLUSION:SVV may act as an angiogenic factor and used for therapeutic angiogenesis in peripheral arterial diseases.
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