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Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF_(165) and angiopoietin-1 gene expression in vitro

查看全文 作　　者：Hao [1]Zhang;Hongyan [2]Dong;Bo [1]Jiang;Zheng [1]Wang;Rui [1]Chen;Zhifeng [2]Zhang;Zhongming [1]Zhang 高影响力作者 机构地区：[1]Institute of Cardiovascular Disease, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu 221002, China;[2]Center of Biological Research, Xuzhou Medical College, Xuzhou, Jiangsu 221002, China.高影响力机构 出　　处：《The Journal of Biomedical Research》索引2011年第25卷第3期,共9页高影响力期刊 基　　金：supported by a grant from the National Natural Science Foundation of China (No.30672081) 摘　　要：Angiogenesis in ischemic tissue is a complex and multi-gene event. In the study, we constructed hypoxic re-sponse elements (HRE) and the Tet-On advanced double-controlled systems and investigated their effects on the expression of hVEGF165 and angiopoietin-1 (Ang-1) genes in rat cardiomyocytes exposed to hypoxia and pharma-cologic induction. We infected neonatal rat cardiomyocytes with recombinant rAAV-rtTA-Rs-M2/rAAV-TRE-Tight-Ang-1 and rAAV-9HRE- hVEGF165. Our results indicated that the viral titer was 1×1012 vg /mL and the viral purity exceeded 98%. hVEGF165 expression was induced by hypoxia, but not by normoxia (P < 0.001). Ang-1 expression was evident under doxycycline induction, but undetectable without doxycycline induction (P < 0.001). Immunofluorescence staining showed that positively stained hVEGF165 and Ang-1 protein appeared only under both hypoxia and doxycycline induction. We demonstrate here that HRE and the recombinant Tet-On advanced double gene-controlled systems sensitively regulate the expression of hVEGF165 and Ang-1 genes in an altered oxygen environment and under pharmacological induction in vitro. 关 键 词：血管生成素-1 双控制系统 多基因 缺氧 春节 反应元件 体外 诱导表达
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