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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 investigate the influence of CD74 gene transfection conditions and CD74 gene transfection on cell characteristics and function in HUVEC and lay the foundation for further study of biological therapy based on CD74 expression. Methods (1) The expression of functional-related genes (HLA-A, HLA-DR and IFNGR) in ECV304 cells before and after transfection with pCMV-CD74: Real-time RT-PCR was used to compare the function-related genes of umbilical vein endothelial cells before and after transfection (2) Real-time RT-PCR analysis software was used to analyze the experimental data. Results The results of Real-time RT-PCR showed that the HLA-A △ Ct values of ECV304 cells before and after transfection were 9.4881 and 12.1097, respectively, and the value of 6.1543 was greater than 2, indicating that HLA-A expression was significantly different before and after transfection; IFNGR △ Ct values were 4.9828 and 5.5228, respectively. There was no significant difference between the two groups (1.4539 and less than 2. Ct was 1.4539). The HLA-DR △ Ct values were 14.1098 and 14.7924, respectively. No significant difference before and after. CONCLUSION: Human umbilical vein endothelial cells (ECV304) express HLA-A, HLA-DR, IFNGR and CD74 mRNA. The transfection of pCMV-CD74 can increase the expression of CD74 gene and membrane molecule of ECV304 cells and increase the expression of HLA- DR and IFNGR expression, in order to further explore the CD74 gene and endothelial cell function to provide experimental data.