Expression of KDRn3 Protein in Human Umbilical Vein Endothelial Cells and Inhibitory Effect of Expressed Product on Cell Proliferation
Ying Pei
Abstract
Ying Pei
Abstract
Objective To observe the expression of KDRn3 protein in human umbilical vein endothelial cells (HUVECs) as well as the inhibitory effect of expressed product on the proliferation of HUVECs. Methods Plasmid pEGFP-N1 / KDRn3 constructed previously was amplified and identified, then transfected to HUVECs in mediation of liposome. Culture the transfected HUVECs for a certain time, then observe the expression of green fluorescent protein by fluorescent microscopy, determine the KDRn3 content in culture supernatant by ELISA and the proliferation level of HUVECs by MTT method. Results Green fluorescence was observed in the HUVECs 48 h after transfection with pEGFP-N1 / KDRn3. However, both the number of HUVECs showing green fluorescence and the fluorescence strength increased 72 h after transfection. The KDRn3 contents in cell culture supernatant 24, 48 and 72 h after transfection were significantly higher than those before transfection. The proliferations of HUVECs 48 and 72 h after transfection with pEGFP-N1 / KDRn3 were significantly inhibited as compared with those of control. Conclusion KDRn3 protein was expressed in HUVECs and inhibited the proliferation of the latter.
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Objective To observe the expression of KDRn3 protein in human umbilical vein endothelial cells (HUVECs) as well as the inhibitory effect of expressed product on the proliferation of HUVECs. Methods Plasmid pEGFP-N1 / KDRn3 constructed previously was amplified and identified, then transfected to HUVECs in mediation of liposome. Culture the transfected HUVECs for a certain time, then observe the expression of green fluorescent protein by fluorescent microscopy, determine the KDRn3 content in culture supernatant by ELISA and the proliferation level of HUVECs by MTT method. Results Green fluorescence was observed in the HUVECs 48 h after transfection with pEGFP-N1 / KDRn3. However, both the number of HUVECs showing green fluorescence and the fluorescence strength increased 72 h after transfection. The KDRn3 contents in cell culture supernatant 24, 48 and 72 h after transfection were significantly higher than those before transfection. The proliferations of HUVECs 48 and 72 h after transfection with pEGFP-N1 / KDRn3 were significantly inhibited as compared with those of control. Conclusion KDRn3 protein was expressed in HUVECs and inhibited the proliferation of the latter.
Key concepts: Transfection, Umbilical vein, Molecular biology, Human umbilical vein endothelial cell, Cell growth, Green fluorescent protein, Fluorescence microscope, Cell culture