Prandini Marie-Hélène, Dreher Inge, Bouillot Stéphanie, Benkerri Souhila, Moll Thomas, Huber Philippe
Department of Cellular Responses and Dynamics, CEA-INSERM-Joseph Fourier University EMI 0219, CEA-Grenoble, France.
Oncogene. 2005 Apr 21;24(18):2992-3001. doi: 10.1038/sj.onc.1208483.
Vascular endothelial (VE)-cadherin is exclusively expressed at interendothelial junctions of normal and tumour vessels. In this report, we characterized the transcriptional activity of the human VE-cadherin promoter. Transient transfection assays revealed that sequences at positions --1135/-744 and -166/-5 base pairs are critical for promoter activity in endothelial cells. We show that specific sequences in the proximal region interact with Ets and Sp1 family members. Transgenic mice were created and the human VE-cadherin promoter was able to confer correct temporal and spatial expression on the LacZ gene in embryos. In adults, the transgene was specifically and strongly expressed in the lung, heart, ovary, spleen and kidney glomeruli, whereas expression was weak or absent in the vasculature of other organs, including the brain, thymus, liver and skeletal muscle. Neovessels in tumour grafts and Matrigel implants harboured strong stainings, indicating that promoter activity is enhanced in angiogenic situations. Furthermore, Matrigel and transfection assays showed that VE-cadherin promoter is subjected to bFGF induction. Transgene expression was also noticed in extravascular sites of the central nervous system, suggesting that silencer elements may be located elsewhere in the gene. These results are a first step towards addressing the organ- and tumour-specific regulation of the VE-cadherin gene.
血管内皮(VE)-钙黏蛋白仅在正常血管和肿瘤血管的内皮细胞连接处表达。在本报告中,我们对人VE-钙黏蛋白启动子的转录活性进行了表征。瞬时转染分析表明,位于-1135/-744和-166/-5碱基对位置的序列对于内皮细胞中的启动子活性至关重要。我们发现近端区域的特定序列与Ets和Sp1家族成员相互作用。构建了转基因小鼠,人VE-钙黏蛋白启动子能够在胚胎中赋予LacZ基因正确的时空表达。在成年小鼠中,转基因在肺、心脏、卵巢、脾脏和肾小球中特异性且强烈表达,而在包括脑、胸腺、肝脏和骨骼肌在内的其他器官的脉管系统中表达较弱或不表达。肿瘤移植物和基质胶植入物中的新生血管有强烈染色,表明在血管生成情况下启动子活性增强。此外,基质胶和转染分析表明VE-钙黏蛋白启动子受碱性成纤维细胞生长因子(bFGF)诱导。在中枢神经系统的血管外部位也观察到转基因表达,这表明沉默元件可能位于基因的其他位置。这些结果是朝着解决VE-钙黏蛋白基因的器官和肿瘤特异性调控迈出的第一步。