Hypoxic regulation of endothelial  glyceraldehyde-3-phosphate dehydrogenase

Graven, Krista K.; McDonald, Robert J.; Farber, Harrison W.

doi:10.1152/ajpcell.1998.274.2.c347

Cited by 138 publications

(115 citation statements)

References 35 publications

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“…To analyze transcriptional control, run-off transcription assays were performed that showed a 2.7-fold up-regulation of VPF/VEGF mRNA transcription by hypoxia in M21 cells compared with the 36B4 control, which was unaffected (Figure 2A). In comparison, GAPDH showed a 3-fold and Glut-1 a 2.5-fold transcription up-regulation, consistent with the increased hypoxic expression of both of these genes (Graven et al, 1994;Ebert et al, 1995). The level of increased gene transcription, however, only partially accounted for the total increase in VPF/VEGF mRNA expression observed by Northern blot analysis, suggesting an additional effect of hypoxia on mRNA stability.…”

Section: Hypoxia-induced Vpf/vegf Mrna Expression In M21 Melanoma Celsupporting

confidence: 68%

Identification of a Human VPF/VEGF 3′ Untranslated Region Mediating Hypoxia-induced mRNA Stability

Claffey¹,

Shih²,

Mullen³

et al. 1998

MBoC

154

143

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Hypoxia is a prominent feature of malignant tumors that are characterized by angiogenesis and vascular hyperpermeability. Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) has been shown to be up-regulated in the vicinity of necrotic tumor areas, and hypoxia potently induces VPF/VEGF expression in several tumor cell lines in vitro. Here we report that hypoxia-induced VPF/VEGF expression is mediated by increased transcription and mRNA stability in human M21 melanoma cells. RNAbinding/electrophoretic mobility shift assays identified a single 125-bp AU-rich element in the 3Ј untranslated region that formed hypoxia-inducible RNA-protein complexes. Hypoxia-induced expression of chimeric luciferase reporter constructs containing this 125-bp AU-rich hypoxia stability region were significantly higher than constructs containing an adjacent 3Ј untranslated region element without RNA-binding activity. Using UV-cross-linking studies, we have identified a series of hypoxia-induced proteins of 90/88 kDa, 72 kDa, 60 kDa, 56 kDa, and 46 kDa that bound to the hypoxia stability region element. The 90/88-kDa and 60-kDa species were specifically competed by excess hypoxia stability region RNA. Thus, increased VPF/VEGF mRNA stability induced by hypoxia is mediated, at least in part, by specific interactions between a defined mRNA stability sequence in the 3Ј untranslated region and distinct mRNA-binding proteins in human tumor cells. INTRODUCTIONVascular permeability factor/vascular endothelial growth factor (VPF/VEGF) 1 is a potent activator of microvascular permeability in vivo and an endothelial cell-specific mitogen in vitro Ferrara and Henzel, 1989;Gospodarowicz et al., 1989;Keck et al., 1989;Leung et al., 1989;Levy et al., 1989;Conn et al., 1990;Senger et al., 1990; Ferrara et al., 1991a,b;Dvorak et al., 1992). VPF/VEGF expression has been closely associated with the pathological angiogenesis observed in malignant tumors (Plate et al., 1992; Brown et al., 1993a,b;Weindel et al., 1994; Brown et al., 1995a,b), diabetic retinopathy (Adamis et al., 1994;Aiello et al., 1994), retinopathy of prematurity Pierce et al., 1995;Stone et al., 1995), rheumatoid arthritis (Fava et al., 1994), and coronary artery disease (Sabri et al., 1991;Ladoux and Frelin, 1993;Hashimoto et al., 1994). Tissue hypoxia is a common feature in many of these diseases and VPF/VEGF expression is dramatically up-regulated in human solid tumors adjacent to sites of focal necrosis (Plate et al., 1992;Shweiki et al., 1992; Brown et al., 1993).Hypoxia-stimulated VPF/VEGF expression has been attributed to increases in both transcriptional and posttranscriptional mechanisms (Ikeda et al., 1995;Stein et al., 1995; Levy et al., 1996a,b). The transcriptional up-regulation of VPF/VEGF under hypoxia appears to be a common mechanism in a multitude of † Corresponding author. 1 Abbreviations used: ARE, adenylate-uridylate-rich region;EMSA, electrophoretic mobility shift assay; GM-CSF, granulocyte macrophage-colony stimulating factor; HSR, hypoxia stab...

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Section: Hypoxia-induced Vpf/vegf Mrna Expression In M21 Melanoma Celsupporting

confidence: 68%

Identification of a Human VPF/VEGF 3′ Untranslated Region Mediating Hypoxia-induced mRNA Stability

Claffey¹,

Shih²,

Mullen³

et al. 1998

MBoC

154

143

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“…The reference gene selection was addressed in our studies as it is reported that hypoxia is able to alter the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 37 which has been commonly used as a reference gene in many chondrocyte quantitative PCR analyses. A reference gene selection study was therefore performed.…”

Section: Discussionmentioning

confidence: 99%

Effects of hypoxia on glucose transport in primary equine chondrocytes in vitro and evidence of reduced GLUT1 gene expression in pathologic cartilage in vivo

Peansukmanee¹,

Vaughan-Thomas²,

Carter³

et al. 2009

Journal Orthopaedic Research

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Articular chondrocytes exist in an environment lacking in oxygen and nutrients due to the avascular nature of cartilage. The main source of metabolic energy is glucose, which is taken up by glucose transporters (GLUTs). In diseased joints, oxygen tensions and glucose availability alter as a result of inflammation and changes in vascularisation. Accordingly, in this study we examined the effects of hypoxia and the hypoxia mimetic cobalt chloride (CoCl 2 ) on glucose transport in equine chondrocytes and compared expression of the hypoxia responsive GLUT1 gene in normal and diseased cartilage. Monolayers of equine chondrocytes were exposed to 20% O 2 , 1% O 2 , CoCl 2 (75 mM), or a combination of 1% O 2 and CoCl 2 . Glucose uptake was measured using 2-deoxy-D-[2,6-3 H] glucose. GLUT1 protein and mRNA expression were determined by FACS analysis and qPCR, respectively. GLUT1 mRNA expression in normal and diseased cartilage was analyzed using explants derived from normal, OA, and OCD cartilage. Chondrocytes under hypoxic conditions exhibited a significantly increased glucose uptake as well as upregulated GLUT1 protein expression. GLUT1 mRNA expression significantly increased in combined hypoxia-CoCl 2 treatment. Analysis of clinical samples indicated a significant reduction in GLUT1 mRNA in OA samples. In OCD samples GLUT1 expression also decreased but did not reach statistical significance. The increase in glucose uptake and GLUT1 expression under hypoxic conditions confirms that hypoxia alters the metabolic requirements of chondrocytes. The altered GLUT1 mRNA expression in diseased cartilage with significance in OA suggests that reduced GLUT1 may contribute to the failure of OA cartilage repair. ß

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“…A possible explanation for the discrepancy of our data with other studies might be the use of GAPDH as a reference gene in the quantitative RT-PCR assay. GAPDH was demonstrated as one of several genes which are up-regulated in breast ductal carcinoma compared to histologically normal tissue (Bini et al, 1997) and a significant correlation of VEGF mRNA and GAPDH mRNA expression was found in breast carcinomas (Scott et al, 1998b), possibly because both genes are regulated by hypoxia (Graven et al, 1998). Potentially, an increased number of VEGF mRNA transcripts parallel to an up-regulation of GAPDH in carcinomatous tissue might have caused the similar VEGF/GAPDH ratios in malignant and benign breast tissue allowing only separate comparisons in our study for benign and malignant tissue (Figure 2).…”

Section: Discussionmentioning

confidence: 99%

Vascular endothelial growth factor A (VEGF-A) mRNA expression levels decrease after menopause in normal breast tissue but not in breast cancer lesions

Greb¹,

Maier²,

Wallwiener³

et al. 1999

Br J Cancer

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SummaryWe hypothesized that the regulation of microvascular functions and angiogenesis in breast tissue, a well known target of ovarian steroid action, is dependent on the hormonal exposure of the breast. Relative expression levels of VEGF-A (vascular endothelial growth factor A), a putative key regulator of angiogenesis in breast cancer, were analysed in the tumour and the adjacent non-neoplastic breast tissue of 19 breast cancer patients by quantitative reverse transcriptase polymerase chain reaction. In non-neoplastic breast specimens the expression levels of all detected VEGF-A-isoforms (189, 165, 121) were significantly higher in premenopausal compared to post-menopausal women (P = 0.02) and were inversely correlated with the patient's age (P = 0.006). In contrast, in cancerous tissues menopausal status had no influence on VEGF-A-expression levels. Benign and malignant tissues exhibited a similar expression pattern of VEGF-A-isoforms relative to each other. Thus, the regulation of the vasculature in normal breast tissue, as opposed to breast cancer tissue, appears to be hormonally dependent. Endogenous and therapeutically used hormonal steroids might, therefore, cause clinically relevant changes of the angiogenic phenotype of the human breast.

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Hypoxic regulation of endothelial glyceraldehyde-3-phosphate dehydrogenase

Cited by 138 publications

References 35 publications

Identification of a Human VPF/VEGF 3′ Untranslated Region Mediating Hypoxia-induced mRNA Stability

Identification of a Human VPF/VEGF 3′ Untranslated Region Mediating Hypoxia-induced mRNA Stability

Effects of hypoxia on glucose transport in primary equine chondrocytes in vitro and evidence of reduced GLUT1 gene expression in pathologic cartilage in vivo

Vascular endothelial growth factor A (VEGF-A) mRNA expression levels decrease after menopause in normal breast tissue but not in breast cancer lesions

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