CD74基因转染对内皮细胞特性的影响

Impact of gene transfection on the expression of vein endothelial cell

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。

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.