Identification of hypoxia in cells and tissues of epigastric 9L rat glioma using EF5 [2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl) acetamide]

Evans, Sydney M.; Joiner, B.; Jenkins, W. Timothy; Laughlin, Kristine M.; Lord, Edith M.; Koch, Cameron J.

doi:10.1038/bjc.1995.427

Cited by 103 publications

(47 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Intracellular metabolism of EF5 leads to its covalent binding with several molecules. These bound adducts can be detected with specific antibodies (Evans et al, 1995). Our results showed EF5 binding in the brain at 1 hr but not at 2 d of recovery from cardiac arrest.…”

Section: Discussionmentioning

confidence: 74%

“…Once animals were killed, the brains were quickly removed and frozen at Ϫ70°C. To detect EF5 adducts, 10 m coronal sections were incubated with an EL K3-51 monoclonal C y3-conjugated antibody (provided by S. Evans, University of Pennsylvania, Philadelphia, PA) according to the procedure described by Evans et al (1995).…”

Section: Induction Of Transient Global Ischemia By Cardiac Arrest Andmentioning

confidence: 99%

See 1 more Smart Citation

Activation of Hypoxia-Inducible Factor-1 in the Rat Cerebral Cortex after Transient Global Ischemia: Potential Role of Insulin-Like Growth Factor-1

Chávez¹,

2002

View full text Add to dashboard Cite

Hypoxia-inducible factor-1 (HIF-1) is a transcription factor that regulates the adaptive response to hypoxia in mammalian cells. It consists of a regulatory subunit HIF-1␣, which accumulates under hypoxic conditions, and a constitutively expressed subunit HIF-1␤.In this study we analyzed HIF-1␣ expression in the rat cerebral cortex after transient global ischemia induced by cardiac arrest and resuscitation. Our results showed that HIF-1␣ accumulates as early as 1 hr of recovery and persists for at least 7 d.In addition, the expression of HIF-1 target genes, erythropoietin and Glut-1, were induced at 12 hr to 7d of recovery. A logical explanation for HIF-1␣ accumulation might be that the brain remained hypoxic for prolonged periods after resuscitation. By using the hypoxic marker 2-(2-nitroimidazole-1[H]-y1)-N- (2,2,3,3,3-pentafluoropropyl)-acetamide (EF5), we showed that the brain is hypoxic during the first hours of recovery from cardiac arrest, but the tissue is no longer hypoxic at 2 d. Thus, the initial ischemic episode must have activated other nonhypoxic mechanisms that maintain prolonged HIF-1␣ accumulation. One such mechanism might be initiated by insulin-like growth factor-1 (IGF-1). Our results showed that IGF-1 expression was upregulated after cardiac arrest and resuscitation. In addition, we showed that IGF-1 was able to induce HIF-1␣ in pheochromocytoma cells and cultured neurons as well as in the brain of rats that received intracerebroventricular and systemic IGF-1 infusion. Moreover, infusion of a selective IGF-1 receptor antagonist abrogates HIF-1␣ accumulation after cardiac arrest and resuscitation. Our study suggest that activation of HIF-1 might be part of the mechanism by which IGF-1 promotes cell survival after cerebral ischemia.

show abstract

Section: Discussionmentioning

confidence: 74%

Section: Induction Of Transient Global Ischemia By Cardiac Arrest Andmentioning

confidence: 99%

Activation of Hypoxia-Inducible Factor-1 in the Rat Cerebral Cortex after Transient Global Ischemia: Potential Role of Insulin-Like Growth Factor-1

Chávez¹,

2002

View full text Add to dashboard Cite

show abstract

“…To identify hypoxic regions of the primary tumor, mice received an injection of the hypoxia marker drug EF5 [2-(2-nitro-1H-imidazole-1-yl)-N-(2,2,3,3,3-pentafluoropropyl) acetamide], obtained from Dr. Cameron Koch (University of Pennsylvania), 3 h before tumor excision (38). To achieve optimal drug distribution and tumor staining, each mouse received 5 l/g i.v.…”

Section: Methodsmentioning

confidence: 99%

Acute Hypoxia Enhances Spontaneous Lymph Node Metastasis in an Orthotopic Murine Model of Human Cervical Carcinoma

2004

View full text Add to dashboard Cite

An orthotopic mouse model of cervical carcinoma has been used to investigate the relationship between acute (cyclic) hypoxia and spontaneous lymph node metastasis in vivo. The human cervical carcinoma cell line ME-180 was stably transfected to express the fluorescent protein DsRed2, which allowed the in vivo optical monitoring of tumor growth and metastasis by fluorescent microscopy. The surgically implanted primary tumors metastasize initially to local lymph nodes and later to lung, a pattern consistent with the clinical course of the disease. The effect of acute hypoxia on the growth and spread of these tumors was examined by exposing tumor-bearing mice to treatment consisting of exposure to 12 cycles of 10 min 7% O 2 followed by 10 min air (total 4 h) daily during tumor growth. After 21 days, the tumors were excised, lymph node and lung metastases were quantified, and the hypoxic fraction and relative vascular area of the primary tumors were assessed by immunohistochemical staining for the hypoxic marker drug EF5 [2-(2-nitro-1H-imidazole-1-yl)-N-(2,2,3,3,3-pentafluoropropyl) acetamide] and the vascular marker CD31, respectively. In untreated mice, the primary tumor size was directly correlated with lymph node metastatic burden. The acute hypoxia treatment resulted in a significant decrease in the size of the primary tumors at the time of excision. However, the mice in the acute hypoxia group had an increased number of positive lymph nodes (2-4) as compared with control mice (1-3). Lung metastasis was not affected. The acute hypoxia treatment also decreased the relative vascular area in the primary tumors but did not affect the hypoxic fraction. These results suggest that fluctuating oxygenation in cervical carcinoma tumors may reduce tumor growth rate, but it may also enhance the ability of tumor cells to metastasize to local lymph nodes.

show abstract

“…The use of macrophage-specific antibodies, in conjunction with such hypoxia-specific markers as piminadazole 64 (Fig. 1) or EF5, 65 should allow the microdissection of TAMs from hypoxic versus well-oxygenated areas of tumor sections using laser capture. Gene expression in the RNA extracted from these samples could then be analyzed by cDNA microarrays.…”

Section: Tam Gene Expression In Response To Tumor Hypoxiamentioning

confidence: 99%

Macrophage migration and gene expression in response to tumor hypoxia

Murdoch

Lewis

2005

Intl Journal of Cancer

184

143

View full text Add to dashboard Cite

Monocytes are recruited into tumors from the circulation along defined chemotactic gradients and they then differentiate into tumor-associated macrophages (TAMs). Recent evidence has shown that large numbers of TAMs are attracted to and retained in avascular and necrotic areas, where they are exposed to tumor hypoxia. At these sites, TAMs appear to undergo marked phenotypic changes with activation of hypoxia-inducible transcription factors, dramatically upregulating the expression of a large number of genes encoding mitogenic, proangiogenic and prometastatic cytokines and enzymes. As a consequence, high TAMs density has been correlated with increased tumor growth and angiogenesis in various tumor types. Since hypoxia is a hallmark feature of malignant tumors and hypoxic tumor cells are relatively resistant to radio-and chemotherapy, these areas have become a target for novel forms of anticancer therapy. These include hypoxiatargeted gene therapy in which macrophages are armed with therapeutic genes that are activated by hypoxia-responsive promoter elements. This restricts transgene expression to hypoxic areas, where the gene product is then released and acts on neighboring hypoxic tumor cells or proliferating blood vessels. In this way, the responses of macrophages to tumor hypoxia can be exploited to deliver potent antitumor agents to these poorly vascularized, and thus largely inaccessible, areas of tumors. ' 2005 Wiley-Liss, Inc.

show abstract

Identification of hypoxia in cells and tissues of epigastric 9L rat glioma using EF5 [2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl) acetamide]

Abstract: Identification of hypoxia in cells and tissues of epigastric 9L rat glioma using EF5 [2-(2-nitro-lH-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)

Cited by 103 publications

References 29 publications

Activation of Hypoxia-Inducible Factor-1 in the Rat Cerebral Cortex after Transient Global Ischemia: Potential Role of Insulin-Like Growth Factor-1

Activation of Hypoxia-Inducible Factor-1 in the Rat Cerebral Cortex after Transient Global Ischemia: Potential Role of Insulin-Like Growth Factor-1

Acute Hypoxia Enhances Spontaneous Lymph Node Metastasis in an Orthotopic Murine Model of Human Cervical Carcinoma

Macrophage migration and gene expression in response to tumor hypoxia

Contact Info

Product

Resources

About