4.6

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2.2

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  • ISSN 1674-8301
  • CN 32-1810/R
Volume 25 Issue 3
Apr.  2011
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Hao Zhang, Hongyan Dong, Bo Jiang, Zheng Wang, Rui Chen, Zhifeng Zhang, Zhongming Zhang. Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF165 and angiopoietin-1 gene expression in vitro[J]. The Journal of Biomedical Research, 2011, 25(3): 204-212. DOI: 10.1016/S1674-8301(11)60027-4
Citation: Hao Zhang, Hongyan Dong, Bo Jiang, Zheng Wang, Rui Chen, Zhifeng Zhang, Zhongming Zhang. Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF165 and angiopoietin-1 gene expression in vitro[J]. The Journal of Biomedical Research, 2011, 25(3): 204-212. DOI: 10.1016/S1674-8301(11)60027-4
shu

Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF165 and angiopoietin-1 gene expression in vitro

  • 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

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This study was supported by a grant from the National Natural Sci-ence Foundation of China (No.30672081)

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    • Abstract
  • 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.
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