Selection and Analysis of a Mutant Cell Line Defective in the Hypoxia-inducible Factor-1 α-Subunit (HIF-1α)

Wood, S M; Wiesener, Michael S.; Yeates, K. M.; Okada, Nobuhiko; Pugh, Christopher W.; Maxwell, PH; Ratcliffe, Peter J.

doi:10.1074/jbc.273.14.8360

Cited by 176 publications

(75 citation statements)

References 37 publications

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“…Similar results were obtained for EPO-HRE marker induction in the C15 cells. In contrast, and in accordance with previously published data (Dachs et al, 1997;Wood et al, 1998), anoxia resulted in similar induction levels of both the EPO and PGK-1-HRE constructs in the C4.5 cells (5.0+1.0 and 4.6+2.9-fold respectively) and a sixfold induction of PGK-1-HRE mediated expression in the 9-3C cells.…”

Section: Resultssupporting

confidence: 48%

“…This is in complete agreement with the observations of Ryan et al (2000), who observed no di erence in the vessel density of HIF-1 wt versus HIF-1a de®cient tumours, despite clear di erences in the expression of vascular endothelial growth factor (VEGF), commonly regarded as the key angiogenic factor in tumorigenesis. None of the CHO-derived strains used in the present study induce VEGF expression when exposed to hypoxia in vitro (Wood et al, 1998). Although this may be attributable in part to the lack of HIF-2a in these cell lines, breast carcinoma cell lines lacking HIF-2a have been previously shown to express VEGF (Blancher et al, 2000), suggesting complexities surrounding VEGF expression in di erent cell types.…”

Section: Discussionmentioning

confidence: 61%

“…The CHO derivatives C4.5 and C15, and the human HT1080 ®brosarcoma derived cell lines, 9-3C and 2-C4, all express wt HIF-1. The CHO Ka13.5 strain was derived from C4.5 following drug-induced mutagenesis (Wood et al, 1998), and lacks HIF-1a expression. The C15 cells have been exposed to the same mutagenic procedure as the Ka13.5 strain, and were therefore used as an additional control to the parental C4.5 cells.…”

Section: Resultsmentioning

confidence: 99%

“…The CHO-K1 derivative strain, C4.5, contains two HRE-regulated expression constructs: murine PGK-1-HRE driving the expression of human CD-2 and murine EPO-HRE driving human E-sel expression. The HIF-1a-de®cient Ka13.5 strain was cloned from a population of C4.5 cells following drug-induced mutagenesis (Wood et al, 1998). As a control for the e ects of the mutagenesis procedure, an additional drug-treated CHO-K1 strain (C15) was used, which contains the same EPO-HRE construct as the C4.5 and Ka13.5 cells.…”

Section: Cell Lines and Cell Culturementioning

confidence: 99%

“…We used HIF-1 wild-type (wt) and HIF-1a de®cient Chinese hamster ovary (CHO) cells (Wood et al, 1998) transfected with marker constructs for HRE mediated transcription to evaluate the e ect of cyanide and associated alterations in mitochondrial redox on HIF-1 transactivation. Under the treatment conditions used, there is a greater reliance on the exogenous glucose supply to fuel energy metabolism via glycolysis.…”

Section: Introductionmentioning

confidence: 99%

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A protective role for HIF-1 in response to redox manipulation and glucose deprivation: implications for tumorigenesis

et al. 2002

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We have investigated the role of HIF-1 in the cellular response to redox modulation via the inhibition of oxidative phosphorylation. We demonstrate that manipulation of redox in air, achieved by inhibiting cytochrome oxidase with cyanide, induces HIF-1 mediated transcription in wild-type CHO and HT1080 human tumour cells but not in CHO cells de®cient in the oxygen responsive, HIF-1a sub-unit of HIF-1. Hypoglycaemia attenuates cyanide-mediated transcription in nontransformed HIF-1 wild-type CHO cells but not the human tumour derived cell line. Cells lacking either HIF1a, or the second composite sub-unit of HIF-1, HIF-1b, were markedly more sensitive to the combined stress of perturbed redox and hypoglycaemia than wild-type cells. As such conditions together with hypoxia are prevalent in tumours, these data suggest that HIF-1 may have a protective role in adaptation to the tumour microenvironment. In support of this we demonstrate that HIF-1a de®cient cells are less tumorigenic than wild-type cells. They showed a reduced growth rate when grown as xenografts in nude mice. This was not related to vascular parameters that were identical to those found in HIF-1 wild-type tumours. The HIF-1 de®cient tumours lacked focal expression of Glut-1 in hypoxic tumour regions. Compromized glucose uptake and metabolic adaptation to the tumour micro-environment may form the basis of the reduced tumorigenecity associated with these cells.

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Section: Resultssupporting

confidence: 48%

Section: Discussionmentioning

confidence: 61%

Section: Resultsmentioning

confidence: 99%

Section: Cell Lines and Cell Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A protective role for HIF-1 in response to redox manipulation and glucose deprivation: implications for tumorigenesis

et al. 2002

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Role of hypoxia-inducible factor-1 in hypoxia-induced ischemic tolerance in neonatal rat brain

et al. 2000

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Hypoxia‐inducible factor‐1 (HIF‐1) is a heterodimer composed of HIF‐1α and HIF‐1β protein subunits. This transcription factor is essential for the activation of hypoxia‐inducible genes like erythropoietin, some glucose transporters, the glycolytic enzymes, and vascular endothelial growth factor. Because HIF‐1 activation may promote cell survival in hypoxic tissues, we studied the effect of hypoxic preconditioning on HIF‐1 expression in neonatal rat brain. Hypoxic preconditioning (8% O2 for 3 hours), a treatment known to protect the newborn rat brain against hypoxic‐ischemic injury, markedly increased HIF‐1α and HIF‐1β expression. To support the role of HIF‐1 in protective preconditioning, we also studied the effect of two other known HIF‐1 inducers, cobalt chloride (CoCl2) and desferrioxamine (DFX), on HIF‐1 expression and neuroprotection in newborn brain. HIF‐1α and HIF‐1β protein levels were markedly increased after intraperitoneal injection of CoCl2 (60 mg/kg) and moderately increased after intraperitoneal injection of DFX (200 mg/kg) 1 to 3 hours after the injections. Preconditioning with CoCl2 or DFX 24 hours before hypoxia‐ischemia afforded 75 and 56% brain protection, respectively, compared with that in vehicle‐injected littermate controls. Thus, HIF‐1 activation could contribute to protective brain preconditioning, which could be used in high‐risk deliveries and other clinical situations. Ann Neurol 2000;48:285–296

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Physiologic responses to hypoxia and implications for hypoxia‐inducible factors in the pathogenesis of rheumatoid arthritis

Distler¹,

Wenger²,

Gassmann³

et al. 2004

Arthritis & Rheumatism

100

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This article describes the chemical modification of polyethylene terephthalate (PET) with a variety of compounds containing reactive glycidyl group(s). Four different modifiers, namely, diglycidyl ether of bisphenol‐A (DGEBA), N,N′‐bis[3(carbo‐2′,3′‐epoxypropoxy) phenyl] pyromellitimide (BGPM), triglycidyl glycerol (TGG), and triglycidyl isocyanurate (TGIC) were compared for their reactivity toward PET in the melt phase. It was found that the presence of tertiary nitrogen in the structure of the epoxide modifiers plays the role of in‐built catalyst for their reaction with the end groups of PET. TGIC as a modifier was selected for the detailed investigation of the simultaneously occurring degradation and chain extension/branching reactions in a batch‐melt mixer. The reactions were followed by torque changes, analyzing the products for residual carboxyl content, and by determining insoluble content. It is shown that the rate of the reactive modification of PET melt by TGIC depends upon stoichiometry, temperature, rate of shear, and the chemical composition and the molecular weight (MW) of the PET resin. In general, the results indicate an increase in melt viscosity and insoluble content, whereas an overall decrease in carboxyl content occurs, as defined by the choice of mixing conditions and stoichiometry. Analysis of the batch kinetic data can be useful to define the process requirements for carrying out the reactive modification in continuous extrusion equipment. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 643–652, 2003

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Selection and Analysis of a Mutant Cell Line Defective in the Hypoxia-inducible Factor-1 α-Subunit (HIF-1α)

Cited by 176 publications

References 37 publications

A protective role for HIF-1 in response to redox manipulation and glucose deprivation: implications for tumorigenesis

A protective role for HIF-1 in response to redox manipulation and glucose deprivation: implications for tumorigenesis

Role of hypoxia-inducible factor-1 in hypoxia-induced ischemic tolerance in neonatal rat brain

Physiologic responses to hypoxia and implications for hypoxia‐inducible factors in the pathogenesis of rheumatoid arthritis

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