title: 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.
authors.affiliation: Institute of Cardiovascular Disease, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu 221002, China;
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.forename: Hongyan
authors.initials: H
authors.lastname: Dong
authors.forename: Bo
authors.initials: B
authors.lastname: Jiang
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Zhang
authors.forename: Zhongming
authors.initials: Z
authors.lastname: Zhang
keywords: angiopoietin-1; cardiomyocyte; doxycycline; gene control; hypoxia; vascular endothelial growth factor
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