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Hypoxic response elements and Tet-On advanced double-controlled systems regulate hVEGF 165 and angiopoietin-1 gene expression in vitro.

Abstract
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 hVEGF165 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-TRE-Tight-Ang-1 and rAAV-9HRE- hVEGF165. Our results indicated that the viral titer was 1×10(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 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.
AuthorsHao Zhang, Hongyan Dong, Bo Jiang, Zheng Wang, Rui Chen, Zhifeng Zhang, Zhongming Zhang
JournalJournal of biomedical research (J Biomed Res) Vol. 25 Issue 3 Pg. 204-12 (May 2011) ISSN: 1674-8301 [Print] China
PMID23554691 (Publication Type: Journal Article)

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