Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF165 and angiopoietin-1 gene expression in vitro
Hao Zhang
Hongyan Dong
Bo Jiang
Zheng Wang
Rui Chen
Zhifeng Zhang
Zhongming Zhang
摘要：Angiogenesis in ischemic tissue is a complex and multi-gene event. In the study, we constructed hypoxic response elements (HRE) and the Tet-On advanced double-controlled systems and investigated their effects on the expression of hVEGF 165 and angiopoietin-1 (Ang-1) genes in rat cardiomyocytes exposed to hypoxia and pharmacologic induction. We infected neonatal rat cardiomyocytes with recombinant rAAV-rtTA-Rs-M2/rAAV-TRETight-Ang-1 and rAAV-9HRE- hVEGF 165. Our results indicated that the viral titer was 1X 10 12 vg /mL and the viral purity exceeded 98%. hVEGF 165 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 hVEGF 165 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 hVEGF 165 and Ang-1 genes in an altered oxygen environment and under pharmacological induction in vitro.
机标关键词：in vitrogene expressionhVEGF165Tet-OndoxycyclineAng-1response elementsenvironment and
分类号：Q785(基因工程（遗传工程）)
资助基金：This study was supported by a grant from the National Natural Science Foundation of China(30672081)
论文发表日期：2011-01-01
在线出版日期：2025-08-15（本平台首次上网日期，不代表文献的发表时间）
英文信息