Histopathologic and Immunohistochemical Features in Human Skin after Exposure to Nitrogen and Sulfur Mustard

Smith, Kathleen J.; Smith, William J.; Hamilton, Tracey A.; Skelton, Henry G.; Graham, John S.; Okerberg, Carlin V.; Moeller, Robert B.; Hackley, Brennie E.

doi:10.1097/00000372-199802000-00005

Cited by 81 publications

(71 citation statements)

References 11 publications

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“…Dual function transcription factors have been described that switch their function by interaction with different co-activators or co-repressors (40 -42), different neighboring transacting factors (43)(44)(45), or interaction with specific DNA sequences in different locations relative to the transcription start site (46). MDBP/RFX interacts with c-Abl, greatly stimulating its autophosphorylation (47).…”

Section: Discussionmentioning

confidence: 99%

“…The data presented here indicate that MDBP/RFX behaves as transcriptional repressor in the context of methylated collagen ␣2(I) sequence. We have previously demonstrated that in a rat cell line that has lost expression of this gene, there is methylation in the promoter region and that treatment of this cell line with the DNA demethylating agent 5-azacytidine results in the gain of expression of this gene (43). Because abnormal hypermethylation of the 5Ј region of tumor suppressor genes is so frequent in cancer and down-regulation of collagen gene expression has been proposed to contribute to carcinogenesis, abnormal methylation of the 5Ј region of the ␣2(I) gene, including the MDBP/RFX site, might occur in cancers and play a role in down-regulation of the expression of this gene.…”

Section: Discussionmentioning

confidence: 99%

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A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

Sengupta¹,

Ehrlich²,

Smith³

1999

Journal of Biological Chemistry

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DNA methylation inhibits transcription driven by the collagen ␣2(I) promoter and the 5 end of the gene in transient transfection and in vitro transcription assays. DNA-binding proteins in a unique family of ubiquitously expressed proteins, methylated DNA-binding protein (MDBP)/regulatory factor for X box (RFX), form specific complexes with a sequence overlapping the transcription start site of the collagen ␣2(I) gene. Complex formation increased when the CpG site at ؉7 base pairs from the transcription start site was methylated. The identity of the protein was demonstrated by co-migration and cross-competition for a characteristic slowly migrating doublet complex formed on MDBP/RFX recognition sequences and the collagen sequences by band shift assays. A RFX1-specific antibody supershifted the collagen DNA-protein complexes. Furthermore, in vitro translated RFX1 protein formed a specific complex with the collagen sequence that was also supershifted with the RFX1 antibody. MDBP/RFX displayed a higher affinity binding to the collagen sequence if the CpG at ؉7 was mutated in a manner similar to TpG. This same mutation within reporter constructs inhibited transcription in transfection and in vitro transcription assay. These results support the hypothesis that DNA methylationinduced inactivation of collagen ␣2(I) gene transcription is mediated, in part, by increased binding of MDBP/ RFX to the first exon in response to methylation in this region.Type I collagen, the most abundant collagen molecule within the collagen family, normally consists of a heterotrimer of two ␣1(I) chains and one ␣2(I) chain. Synthesis of these genes is often down-regulated upon oncogenic transformation of cells in culture (1-4). We have previously demonstrated down-regulation of the collagen ␣2(I) gene, encoding one of the subunits of Type I collagen in an epithelial-like cell line from rat liver, K16 cells upon their conversion to a tumorigenic line, W8, after treatment with the carcinogen 2-N-(acetoxyacetyl)-aminofluorine (3). The promoter-5Ј region of the ␣2 gene was methylated in the nonexpressing W8 cells and not in the expressing K16 cells (5). Furthermore, reporter gene expression downstream of the 218-bp 1 promoter and the 54-bp 5Ј region of the rat and human collagen ␣2(I) genes was inactivated by in vitro DNA methylation in transient transfection experiments, whereas an analogous expression plasmid with the SV40 early promoter/ enhancer driving expression was not (6). We also demonstrated that a minimal collagen ␣2(I) promoter containing the preinitiation region (Ϫ41 to ϩ54) driving expression of the luciferase reporter gene was inactivated by DNA methylation (7). The inhibition of reporter gene expression was attributable to CpG methylation, specifically of collagen ␣2(I) sequences. However, all the methylation sites were located in the first exon, not in the promoter. DNA methylation in the promoter and 5Ј region of genes often correlates with decreased transcription of vertebrate genes, and many studies indicate that this methyla...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

Sengupta¹,

Ehrlich²,

Smith³

1999

Journal of Biological Chemistry

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“…Sulfur mustard (bis(2-chloroethyl)sulfide, HD) is a chemical warfare agent that penetrates the skin rapidly and causes extensive blistering after a latent period of several hours (1)(2)(3)(4)(5)(6).…”

Section: Introductionmentioning

confidence: 99%

“…This study is divided into two sections: (1) a time course study and (2) a compound evaluation study. The specific aim of the time course study was to determine whether MMP and MMP substrate gene expression levels are altered over time (6,12,24, and 72 h) in mouse ear skin topically exposed to liquid HD. Gene expression profiles support the identification of specific targets for therapeutic countermeasures against HD-induced skin injury and provide additional biochemical markers for understanding HD toxicity.…”

Section: Introductionmentioning

confidence: 99%

Use of Epidermolysis Bullosa Biomarkers in Models of Vesicant Injury

Gerecke¹

2006

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“…Yet at this time, no single drug or combination of known protective agents appear to afford enough protection to be considered a sulfur mustard therapeutic agent (10).…”

Section: Introductionmentioning

confidence: 99%

A Double-Label Technique that Monitors Sulfur Mustard Damage to Nuclei and Mitochondria of Normal Human Epidermal Keratinocytes In Vitro

Cook

Buskirk

1997

Toxicol Pathol

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Sulfur mustard and 2-chloro ethyl ethyl sulfide (CEES, a sulfur mustard analog) is known to have immediate (minutes), long-term (hours to days), and toxic effects on human skin. Research was directed toward developing a single in vitro assay that might reflect both these short-term and long-term effects of this vesicating agent on normal human epidermal keratinocytes (NHEK) in vitro. Such an assay system would be useful in identifying and developing sulfur mustard therapeutic agents. NHEK were exposed to the monofunctional sulfur mustard analog 2-chloro ethyl ethyl sulfide for a variety of times. The effects of CEES on NHEK nuclei were assessed using the membrane-permeable SYTO nuclear stains, whereas the effects of CEES on NHEK metabolism were determined by using the nontoxic mitochondria dye Alamar blue. CEES enhanced SYTO binding in a concentration-dependent manner to the nucleus immediately subsequent to a 2-hr exposure, whereas CEES had relatively little effect on metabolic activity at this time. Fifteen to 36 hr subsequent to CEES exposure, however, Alamar blue revealed a robust, sulfur mustard-dependent effect on mitochondrial activity. To determine if both these indicator dyes could be used simultaneously, NHEK were exposed to CEES and stained with the SYTO nuclear stain 2 hr subsequent to exposure. This procedure was followed by assay of the same cell cultures with Alamar blue at 36 hr subsequent to initial CEES exposure. The data indicate that this nuclear/mitochondrial double-label technique can be used to monitor the short-and long-term effects of sulfur mustard on the same culture of NHEK.

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Histopathologic and Immunohistochemical Features in Human Skin after Exposure to Nitrogen and Sulfur Mustard

Cited by 81 publications

References 11 publications

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

Use of Epidermolysis Bullosa Biomarkers in Models of Vesicant Injury

A Double-Label Technique that Monitors Sulfur Mustard Damage to Nuclei and Mitochondria of Normal Human Epidermal Keratinocytes In Vitro

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