CD74基因转染对内皮细胞特性的影响

目的探讨CD74基因在HUVEC的转染条件和CD74基因转染对细胞特性及功能的影响,为深入研究基于调节CD74表达的生物治疗打下基础。方法 （1）pCMV-CD74质粒转染前后ECV304细胞功能相关基因（HLA-A、HLA-DR、IFNGR）表达的检测：采用Real-time RT-PCR比较转染前后脐静脉内皮细胞各功能相关基因的表达;（2）应用Real-time RT-PCR分析软件分析实验数据。结果 Real-time RT-PCR结果显示,转染前后ECV304细胞HLA-A△Ct值分别为9.4881和12.1097,2-△△Ct为6.1543大于2,说明转染前后HLA-A表达有显著差异;IFNGR△Ct值分别为4.9828和5.5228,2-△△Ct为1.4539小于2,转染前后无显著差异;HLA-DR△Ct值分别为14.1098和14.7924,2-△△Ct为1.3036小于2,转染前后无显著差异。结论人脐静脉内皮细胞ECV304表达HLA-A、HLA-DR、IFNGR和CD74mRNA;pCMV-CD74转染可增加ECV304细胞CD74基因和膜分子的表达,增加HLA-A基因的表达,但不增加HLA-DR和IFNGR的表达,为深入探讨CD74基因与内皮细胞功能的相关性提供实验数据。

Impact of gene transfection on the expression of vein endothelial cell

Objective To transfect the CD74 gene to ECV304 cells for observing the impact of transfecting HUVEC on the expression of CD74.Methods The expression of some function-associated genes（HLA-A,HLA-DR and IFNGR）before and after transfected ECV304 by Real-time PCR was detected.Results The △Ct values of HLA-A before and after transfecting CD74 gene to ECV304 were 9.4881 and 12.1097,and 2-△△Ct was 6.1543 showing that the expression of HLA-A was significantly different.The values of IFNGR and HLA-DR were 4.9828,5.5228,the 2-△△Ct was 1.4539,while the HLA-DR and △Ct were 14.1098 and 14.7924,2-△△Ct was 1.3036 recpectively showing that there was no signficant differences in the expressions of IFNGR and HLA-DR before and after transfection.Conclusion Human umbilical vein endothelial cell ECV304 can express HLA-A,HLA-DR,IFNGR and CD74 mRNA.ECV304 can increase the expression of CD74 gene and membrane molecules and it can also increase the expression of HLA-A gene,but it can＇t increase the expression of HLA-DR and IFNGR.