摘要
目的观察KDRn3蛋白在人脐静脉血管内皮细胞中的表达及对其增殖的抑制作用。方法将质粒pEGFP-N1/KDRn3扩增并鉴定后,以脂质体介导转染人脐静脉血管内皮细胞,培养一定时间后,在荧光显微镜下观察绿色荧光蛋白的表达,ELISA检测细胞培养上清中KDRn3的含量,MTT法检测KDRn3蛋白对人脐静脉血管内皮细胞增殖的影响。结果转染后48h,在荧光显微镜下可见pEGFP-N1/KDRn3转染组人脐静脉血管内皮细胞发出绿色荧光,72h发出绿色荧光的细胞增多,强度增强;转染后24、48和72h,细胞培养上清中KDRn3含量与转染前比较均提高,且差异有显著意义;转染后48和72h,pEGFP-N1/KDRn3转染组与对照组比较,细胞增殖有明显的抑制,且差异有显著意义。结论KDRn3蛋白可在人脐静脉血管内皮细胞中表达,并能抑制其增殖。
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.
出处
《中国生物制品学杂志》
CAS
CSCD
2008年第9期768-770,776,共4页
Chinese Journal of Biologicals
基金
长春市科技计划项目(2006104)
关键词
血管内皮生长因子受体
KDRn3蛋白
基因转染
人脐静脉血管内皮细胞
细胞增殖
抑制作用
Vascular endothelial growth factor receptor
KDRn3 protein
Gene transfection
Human umbilical vein endothelial cells （HUVECs）
Cell proliferation
Inhibitory effect
