Effect of 5-bromo-2-deoxyuridine labeled umbilical cord mesenchymal stem cells on cell biology behaviors in vitro

Abstract:

Objective：To observe the ability of cell proliferation，anti-apoptosis，and angiogenesis of rat artery endothelial cells （RAECs） after survivin（SVV） gene transfection. Methods：The SVV was transfected into rat artery endothelial cells（RAECs） by adenovirus. The proliferation rates of RAECs with or without SVV transfection were determined by CCK8 assay .The levels of birc5，caspase-3，bcl-2 and cyclin D before or after transfection were detected by real-time PCR and Western blot. The apoptosis of RAECs was induced by H2O2 and the apoptosis rate in each group was detected by flow cytometry. The angiogenesis was detected by immuno－histochemistry with the RAECs specific marker CD31. Results：The birc5 gene and survivin protein levels were both elevated in SVV over-expressed RAECs determined by RT-qPCR and Western-blot assay（birc5：P=0.006，SVV protein：P=0.000）. Meanwhile，the Caspase-3 protein levels were decreased and the bcl-2，cyclinD levels were increased（Caspase-3：P=0.000；bcl-2：P=0.000；cyclinD：P=0.000）. RAECs transfected with SVV exhibited a significant higher proliferation rate. The microvessel density（MVD） was much higher in RAECs transfected with SVV than in control group. Conclusion：The ability of proliferation，anti-apoptosis and angiogen－esis is significantly elevated in RAECs after SVV transfection.